MENGULAS ARTIKEL

Depan. Penuaan Neurosci, 23 Juli 2013.

| https://doi.org/10.3389/fnagi.2013.00036

Sirtuins: dari regulasi metabolisme penuaan otak

1,2,3 *
Wenzhen Duan
1
Divisi Neurobiologi, Departemen Psikiatri dan Ilmu Perilaku, Johns
Hopkins University School of Medicine, Baltimore, MD, USA
2
Departemen Neuroscience, Johns Hopkins University School of Medicine,
Baltimore, MD, USA
3
Program di Seluler dan Molekuler Kedokteran, Johns Hopkins University
School of Medicine, Baltimore, MD, USA

penuaan otak yang ditandai dengan hilangnya progresif fungsi neurofisiologis

yang sering disertai dengan neurodegeneration terkait usia. pembatasan kalori
telah dikaitkan dengan perpanjangan umur dan pengurangan risiko penyakit
neurodegenerative dalam sistem model eksperimen. Beberapa jalur sinyal telah
ditunjukkan mendasari efek menguntungkan dari pembatasan kalori, di
antaranya keluarga sirtuin telah disarankan untuk memainkan peran
sentral. Pada mamalia, telah ditetapkan bahwa sirtuins mengatur respon
fisiologis untuk metabolisme dan stres, dua faktor utama yang mempengaruhi
proses penuaan. Sirtuins mewakili kelas baru yang menjanjikan dari deacetylases
dilestarikan yang memainkan peran penting dalam mengatur metabolisme dan
penuaan. Ulasan ini berfokus pada pemahaman saat ini tentang hubungan
antara jalur metabolisme yang melibatkan sirtuins dan proses penuaan otak,
dengan fokus pada SIRT1 dan SIRT3. Identifikasi agen terapi mampu memodulasi
ekspresi dan / atau kegiatan dari sirtuins diharapkan dapat memberikan strategi
yang menjanjikan untuk ameliorating neurodegeneration. penyelidikan masa
depan mengenai interaksi bersama dari sirtuins yang berbeda akan membantu
kita memahami lebih lanjut tentang proses penuaan, dan berpotensi mengarah
pada pengembangan pendekatan terapi untuk pengobatan penyakit
neurodegenerative yang berkaitan dengan usia dan promosi penuaan sukses.

pengantar

Otak, mirip dengan organ lain, mengalami penurunan bertahap dalam

metabolisme energi selama penuaan ( Drew dan Leeuwenburgh 2004 ; Navarro
dan Boveris 2007 ; Boveris dan Navarro, 2008 ; Swerdlow, 2011 ). Sejak neuron
memerlukan sejumlah besar energi untuk penembakan potensial aksi,
neurotransmisi, dan proses lainnya, penurunan terkait usia dalam metabolisme
berkontribusi terhadap penurunan kognitif yang terkait dengan penuaan
( Biessels dan Kappelle 2005 ; Boveris dan Navarro, 2008 ). Penuaan juga
merupakan faktor risiko untuk penyakit terkait usia seperti gangguan
neurodegenerative. Penyakit ini dapat terjadi ketika neuron gagal untuk
merespon adaptif untuk penurunan berhubungan dengan usia pada tingkat
metabolisme basal dan dalam tugas-driven energi, seperti koordinasi
neuromuskuler, kinerja kognitif, dan kesadaran lingkungan ( Swerdlow,
2007 ). Dalam dekade terakhir, fungsi sirtuins mamalia, evolusioner nicotinamide
adenin dinukleotida (NAD) -tergantung deacetylases protein / ADP-
ribosyltransferases, telah diteliti secara lebih detail, dan kami sekarang memiliki
pemahaman molekul yang jauh lebih baik dari peran ganda ini keluarga yang
unik enzim bermain di penuaan dan tampaknya setiap proses biologis. Ada
sedikit keraguan bahwa sirtuins telah muncul sebagai modulator kritis tanggapan
adaptif metabolik, dan kegiatan mereka telah dikaitkan dengan beberapa
penyakit, kelainan metabolisme untuk neurodegeneration.

Sirtuins awalnya diidentifikasi sebagai salah satu gen yang mengatur jenis kawin
tunas ragi, Saccharomyces cerevisiae , dan bernama informasi diam regulator 2
(Sir2) dalam organisme yang lebih rendah ( Klar dan Fogel, 1979 ). Menyusul
penerbitan pertama menggambarkan peran ragi Sir2 dalam mempromosikan
umur panjang ( Kaeberlein et al., 1999 ), banyak penyelidikan difokuskan pada
elucidating apakah sirtuins mungkin memainkan peran yang sama pada
organisme lain. Sirtuins telah terbukti untuk mengatur umur pada organisme
yang lebih rendah, termasuk ragi, nematoda, dan lalat buah ( Haigis dan
Guarente 2006 ), meskipun peran mereka dalam cacing dan terbang umur baru-
baru ini diperdebatkan ( Burnett et al, 2011. ; Viswanathan dan Guarente
2011 ). Sebagian besar penelitian ini telah dijelaskan peran kunci untuk SIRT1
dalam mengatur respon metabolik untuk pembatasan kalori (CR; Canto dan
Auwerx, 2009 ), intervensi diet yang kokoh membentang masa hidup di berbagai
spesies. Namun, overekspresi seluruh tubuh dari SIRT1 pada tikus tidak
mempengaruhi rentang hidup ( Herranz et al., 2010 ). Namun demikian, SIRT1
tidak muncul untuk mempromosikan penuaan yang sehat dengan melindungi
terhadap beberapa patologi yang berkaitan dengan usia, seperti memfasilitasi
sensitivitas insulin, mengangkat produksi glukosa, mengurangi stres oksidatif,
potensiasi aktivitas yang diturunkan dari otak faktor neurotropik (BDNF) faktor
transkripsi respon cAMP elemen-mengikat protein (CREB; Guarente dan Franklin,
2011 ).

Mamalia memiliki tujuh sirtuins (SIRT1-7) yang ditemukan di lokasi subselular

yang berbeda, termasuk inti (SIRT1, SIRT6, dan SIRT7), sitosol (SIRT2), dan
mitokondria (SIRT3, SIRT4, dan SIRT5). Sebagian besar penelitian telah dijelaskan
peran kunci untuk SIRT1 dalam mengatur respon metabolik untuk CR ( Canto dan
Auwerx, 2009 ), rejimen diet yang melibatkan berkurang 30-40% asupan kalori
dibandingkan dengan asupan kalori normal, yang mengakibatkan umur
diperpanjang dan mengurangi pengembangan morbiditas dengan penuaan
( Jiang et al, 2000. ; Masoro 2000 ; Sinclair, 2002 ; Koubova dan Guarente
2003 ). Pembatasan kalori adalah satu-satunya intervensi yang secara konsisten
telah ditunjukkan untuk menunda onset, memperlambat perkembangan penyakit
yang berhubungan dengan usia, dan memperpanjang umur pada spesies
berumur pendek, serta berumur panjang primata non-manusia, menunjukkan
bahwa mekanisme yang sama akan operasi pada manusia. Berlebih seluruh
tubuh dari SIRT1 pada tikus tidak mempengaruhi umur ( Herranz et al.,
2010 ). Namun demikian, SIRT1 mempromosikan penuaan yang sehat dengan
mencegah patologi terkait usia ( Guarente dan Franklin, 2011 ). Link lain yang
kuat antara sirtuins mamalia dan efek anti-penuaan dari CR diberikan oleh SIRT3,
yang memediasi pencegahan terkait usia gangguan pendengaran ( Someya et
al., 2010 ). SIRT3 diperlukan untuk pengurangan CR-dimediasi kerusakan
oksidatif pada beberapa jaringan melalui regulasi sistem antioksidan glutathione
( Someya et al., 2010 ).

Dalam ulasan ini, kami fokus pada efek SIRT1 dan SIRT3 pada regulasi
metabolisme dan aktivitas anti-penuaan mereka di otak, dan selanjutnya
membahas pendekatan farmakologis potensi untuk memperbaiki dan mencegah
gangguan neurologis terkait usia dengan menargetkan sirtuins.

SIRT1, metabolisme dan Aging Brain

Distribusi SIRT1 di Otak

Selama embriogenesis mouse, SIRT1 sangat disajikan dalam otak, sumsum

tulang belakang, dan akar dorsal ganglion, dengan ekspresi puncak pada E4.5
( Salminen dan Kaarniranta 2012 ). SIRT1 juga dinyatakan dalam otak orang
dewasa, dengan tingkat tinggi di korteks, hipokampus, serebelum, dan
hipotalamus, dan tingkat rendah dalam materi putih ( Singh, 2004 ). Di antara
berbagai jenis sel otak, SIRT1 didominasi, jika tidak eksklusif, dinyatakan dalam
neuron ( Singh, 2004 ; . Adler et al, 2007 ; Salminen dan Kaarniranta,
2012 ). Satu-satunya pengecualian adalah bahwa SIRT1 ditemukan dalam
mikroglia ketika co-berbudaya dengan neuron ( Schmitz et al., 2004 ). Pada
tingkat subselular, SIRT1 dipandang sebagai protein nuklir ( Chen dan Greene,
2003 ). Namun dilaporkan bahwa SIRT1 memiliki urutan kedua impor nuklir dan
ekspor, dan bahwa SIRT1 hadir dalam fraksi sitosol dari otak tikus, meskipun
fungsi sitosol yang baru mulai akan dijelaskan ( Chen et al, 2005b. ; . Lee et al,
2008 ; Hardie, 2011 ).

SIRT1 menengahi Manfaat Metabolik bawah CR

SIRT1 mengandung 747 asam amino pada manusia, dengan berat molekul
prediksi 81 kDa dan satu diukur dari 120 kDa. Selain histon, SIRT1 juga
deacetylates sejumlah substrat non-histone, termasuk p53 ( Luo et al, 2001. )
Dan Peroksisom proliferator-activated receptor (PPAR) koaktivator-1 (PGC-
1; Nemoto et al, 2005. ), dan FOXO ( . Xiong et al, 2011 ), faktor -light-chain-
enhancer nuklir sel B aktif (NF-kB; . Salminen et al, 2008b ). SIRT1 adalah
menggambar perhatian lebih karena dianggap menjadi salah satu faktor penentu
dalam ekstensi umur disebabkan oleh CR, fenomena yang diamati dalam
organisme filogenetis yang beragam termasuk ragi, cacing, lalat buah, dan
mouse ( Kaeberlein et al, 1999. ; Tissenbaum dan Guarente 2001 ; Howitz et al,
2003. ; Rogina dan Helfand, 2004 ). Peran menguntungkan lebih lanjut didukung
oleh temuan bahwa putatif senyawa SIRT1-mengaktifkan, seperti resveratrol,
juga meningkatkan umur panjang pada beberapa spesies, termasuk ragi ( Howitz
et al., 2003 ), worm ( Wood et al., 2004 ), dan mouse ( Baur et al., 2006 ),
sehingga target anti-penuaan.
Efek dari SIRT1 pada umur panjang bergantung pada aktivitas enzimatik
deasetilasi histon dan non-histon substrat. Sementara deasetilasi histon
mengarah ke interaksi mereka dengan DNA dan konsekuen membungkam gen
( Braunstein et al., 1993 ; Sauve et al, 2006. ; . Dali-Youcef et al, 2007 ), yang
deasetilasi protein non-histon memiliki luas berbagai efek biologis, termasuk
penyesuaian metabolisme, promosi kelangsungan hidup, dan autophagy
( Campisi 2005 ; Dali-Youcef et al, 2007. ; Brooks dan Gu, 2009 ; . Madeo et al,
2010 ). Misalnya, SIRT1 menghambat p53 ( Luo et al., 2001 ), mengurangi efek
pro-apoptosis nya. Hal ini juga menghambat NF-kB ( Yeung et al., 2004 ),
mengurangi efek pro-inflamasi. Sebaliknya, SIRT1 mengaktifkan transkripsi
koaktivator, PGC-1 ( Nemoto et al., 2005 ), yang menyebabkan peningkatan
kadar glukosa, sensitivitas insulin, dan biogenesis mitokondria. Bersama-sama,
dan efek lainnya, memberikan kontribusi pada umur panjang yang ditimbulkan
oleh CR (Gambar 1 ).

GAMBAR 1

GAMBAR 1. Skema diagram dari mekanisme anti-penuaan diaktifkan

oleh SIRT1 dan pembatasan kalori. Pembatasan kalori meregulasi aktivitas
dan tingkat SIRT1, SIRT1 deacetylates substrat, termasuk histone dan molekul
non-histon, dan meningkatkan stabilitas genom, mencegah apoptosis dan
peradangan, dan meningkatkan mitokondria biogenesis, sensitivitas insulin dan
toleransi glukosa. Semua efek ini mengakibatkan umur panjang.

Perubahan metabolisme dan keterlibatan cytoprotective dari CR umumnya

dianggap terjadi pada organ non-saraf, seperti hati, pankreas, otot, dan jaringan
lemak ( Brooks dan Gu, 2009 ; Imai dan Guarente 2010 ). Namun, studi terbaru
menunjukkan bahwa hipotalamus juga dapat menyebabkan efek umur panjang
SIRT1 dan CR melalui koordinasi neurobehavioral dan neuroendokrin perubahan,
termasuk suhu tubuh, nafsu makan, dan aktivitas fisik secara keseluruhan
( Dietrich et al, 2010. ; . Satoh et al, 2010 ). SIRT1 berlimpah dinyatakan di
beberapa daerah di hipotalamus tikus, terutama di arkuata, paraventricular,
ventro- dan inti dorsomedial; dan CR meningkatkan kadar SIRT1 di hipotalamus,
yang meningkatkan suhu tubuh, asupan makanan, dan aktivitas fisik ( Ramadori
et al, 2008. ; . Dietrich et al, 2010 ; Satoh et al, 2010. ). SIRT1 tampaknya
diperlukan untuk perubahan perilaku tersebut, yang dicegah jika SIRT1 yang
tersingkir atau dihambat ( Chen et al, 2005a. ; . Satoh et al, 2010 ). Selain
hipotalamus, SIRT1 juga diungkapkan di daerah lain di otak, termasuk korteks,
striatum, dan hippocampus ( Ramadori et al., 2008 ). Tak lama setelah temuan
ini, peran neuroprotektif SIRT1, telah dilaporkan ( Tang, 2009 ; . Morris et al,
2011 ).
SIRT1 dan Faktor Terlibat dalam CR dan Aging

SIRT1 telah ditemukan untuk menunda penuaan dan meningkatkan umur

panjang dengan mengatur aktivitas protein seluler kunci seperti p53, FOXO dan
Ku70 yang terlibat baik dalam proses apoptosis atau mekanisme perbaikan
sel. SIRT1 sehingga dapat meningkatkan kesehatan dan umur panjang sebagian
dengan baik kematian sel melambat dan / atau dengan meningkatkan
mekanisme perbaikan dalam sel ( Wang et al., 2010 ).

Hal ini telah menjadi semakin jelas bahwa efek bermanfaat dari CR, yang
sebagian karena promosi sirtuins ( Wang et al., 2010 ). Tingkat ekspresi dari
peningkatan SIRT1 pada CR di beberapa binatang pengerat dan jaringan
manusia, termasuk adiposa putih, hati, otot rangka, otak, dan ginjal. Tingkat NAD
telah terbukti meningkat dalam sel hati dalam kondisi CR-seperti, yang pada
gilirannya menginduksi ekspresi SIRT1 ( Rodgers et al., 2005 ). SIRT1 berakhir
mengkonsumsi NAD + sebagai akibat dari aktivitas deacetylase nya,
menghasilkan nicotinamide, inhibitor aktivitas sendiri. NAD + dikenal untuk
melindungi neuron ( Liu et al., 2009 ) dan dengan demikian dengan
meningkatkan tingkat NAD +, CR mungkin aktivitas preserveSIRT1. SIRT1 juga
mengaktifkan PGC1 ( Rodgers et al., 2005 ) yang menghasilkan biogenesis
mitokondria ( Liu et al., 2009 ). Penurunan aktivitas mitokondria dianggap
penyebab di banyak penyakit yang berkaitan dengan usia ( Petersen et al,
2003. ; Singh, 2004 ). CR membangkitkan perbaikan dalam aktivitas mitokondria
mirip dengan SIRT1. Oleh karena itu, adalah mungkin bahwa modulator-molekul
kecil SIRT1 dapat bertindak pada jalur yang sama seperti yang dimodifikasi oleh
CR, dan dengan demikian memiliki potensi untuk mengurangi penyakit yang
berkaitan dengan usia ( Lavu et al., 2008 ).

SIRT1 berinteraksi dengan dan memodulasi faktor kunci lain yang terlibat dalam
penuaan mamalia, seperti NF-kB yang mengontrol peradangan sistemik ringan
bersama dengan penuaan manusia proses-inflamm-penuaan ( Salminen et al.,
2008a ), Target mamalia dari rapamycin (mTOR ; Finley dan Haigis 2009 ), AMP-
activated protein kinase (AMPK; Salminen dan Kaarniranta 2012 ), oleh karena itu
mengontrol proses gin. Proses penuaan melibatkan perubahan peraturan
kekebalan tubuh; NF-kB signaling adalah regulator utama dari sistem kekebalan
tubuh. Penghambatan NF-kB signaling pada tikus berusia dikembalikan
karakteristik jaringan dan ekspresi gen global untuk tikus-tikus muda ( Adler et
al., 2007 ). Fungsi dari kompleks NFkB dapat diatur oleh asetilasi komponen p65
( Schmitz et al., 2004 ). SIRT1 dapat berinteraksi dengan RELA / p65 protein di
NF-kB kompleks dan khusus deacetylates lisin 310, yang telah ditunjukkan untuk
mempotensiasi kapasitas transactivation dari NFkB kompleks ( Chen dan Greene,
2003 ). Beberapa studi telah menunjukkan bahwa SIRT1 adalah inhibitor poten
NF-kB transkripsi ( Yeung et al, 2004. ; Chen et al, 2005b. ). Signaling hubungan
antara SIRT1 dan NF-kB sangat menarik sehubungan dengan penuaan, sebagai
konsekuensi dari rilis rem SIRT1, efisiensi transactivation faktor NF-kB
dipotensiasi, yang membangkitkan aktivasi kekebalan dan inflamm-penuaan.
Proses penuaan juga diatur oleh autophagy. Telah mengidentifikasi jalur sinyal
yang mengatur degradasi autophagic dan SIRT1 adalah regulator ampuh
degradasi autophagic ( Lee et al., 2008 ), SIRT1 dapat berinteraksi dengan dan
deacetylates beberapa komponen di kompleks membentuk autophagosomes,
seperti Atg5, Atg7, dan Atg8 protein ( Lee et al., 2008 ). Ada tumpang tindih yang
jelas antara jaringan pensinyalan yang mengatur kedua penuaan dan
autophagocytosis, yang menekankan pentingnya peran autophagy di mengatur
penuaan dan penyakit degeneratif yang berkaitan dengan usia. Jelaslah bahwa
peningkatan autophagy dapat memperpanjang umur. Kegiatan mTOR ditekan
oleh CR, pengurangan mTOR sinyal adalah mekanisme logika calon manfaat anti-
penuaan CR. Melalui deasetilasi dari berbagai protein yang terlibat dalam proses
autophagy, SIRT1 dapat mengatur proses fisiologis selama penuaan dan
dimoderatori oleh CR ( Haigis dan Guarente 2006 ).

Kontrol yang efisien dari homeostasis metabolisme energi merupakan ciri dari
ditingkatkan umur healthspan dan diperpanjang. AMPK dan SIRT1 jalur sinyal
sangat dilestarikan sensor energi dari peningkatan kadar AMP dan NAD + ,
masing-masing, AMPK signaling terlibat dalam regulasi metabolisme energi
homeostasis ( Hardie, 2011 ). Canto dan Auwerx (2009) menunjukkan bahwa
aktivasi AMPK merangsang aktivitas fungsional SIRT1 dengan meningkatkan
konsentrasi intraseluler dari NAD + . Menariknya, SIRT1 mampu deacetylate
kinase LKB1 yang kemudian meningkat aktivitasnya ( Lan et al., 2008 ). Sejak
LKB1 adalah aktivator hulu AMPK, jalur sinyal ini merangsang aktivasi AMPK. Ini
umpan balik positif antara SIRT1 dan AMPK juga dapat mempotensiasi fungsi
jalur sinyal AMPK-diaktifkan lainnya. Hubungan erat antara AMPK dan SIRT1 bukti
bahwa keseimbangan energi secara efektif mengontrol respon seluler melalui
jaringan sinyal yang terintegrasi. AMPK dapat menghambat aktivitas kompleks
mTOR melalui dua mekanisme yang berbeda, baik dengan langsung fosforilasi
Raptor, komponen peraturan mTORC1, atau oleh fosforilasi protein tuberous
sclerosis 2 (TSC2), yang kemudian menekan aktivitas mTOR ( Jung et al .,
2010 ; Mihaylova dan Shaw, 2011 ). Secara bersama-sama, SIRT1 berinteraksi
dengan jalur sinyal anti-penuaan kunci lainnya sehingga berkontribusi untuk
kontrol umur panjang.

Telah ditetapkan bahwa penuaan merupakan faktor risiko yang diketahui untuk
banyak penyakit neurodegenerative termasuk penyakit Alzheimer (AD), penyakit
Parkinson (PD), Wallerian neurodegeneration, penyakit Huntington (HD), dan
amyotrophic lateral sclerosis (ALS). The pathomechanisms terlibat dalam
gangguan ini melibatkan jalur biokimia umum dan proses, termasuk misfolding
protein, oligomerisasi, dan agregasi, proteolisis, modifikasi pasca-translasi,
disfungsi mitokondria, proses metabolisme normal, dan proinflamasi dan
tanggapan proapoptotik yang kita bahas dalam bagian berikutnya.

SIRT1 dan Age-Associated neurologis Penyakit

degenerasi Wallerian

Degenerasi Wallerian menunjuk pada kematian akson dan degradasi setelah

cedera fokus, diikuti dengan pemecahan selubung mielin. Efek neuroprotektif
SIRT1 terhadap Wallerian degenerasi pertama kali ditemukan pada wlds tikus
transgenik ( Perry et al., 1990 ). Tikus-tikus ini menunjukkan penundaan yang
signifikan dalam degenerasi aksonal setelah cedera fisik atau kimia. Dasar
mekanistik untuk kerusakan akson yang tertunda rupanya terkait dengan wlds
mutan protein chimeric. Telah terbukti bahwa Nicotinamide mononukleotida
adenylyltransferase 1 (NMNAT-1) aktivitas memainkan peran penting dalam
pencegahan kerusakan akson, mengerahkan efek perlindungan melalui aktivasi
SIRT1, sebagai pelindung saraf yang diblokir oleh sirtinol SIRT1 inhibitor atau
siRNA-dimediasi SIRT1 membungkam ( Araki et al, 2004. ; . Sasaki et al,
2009 ; Babetto et al, 2010. ). Peran SIRT1 masih kontroversial, namun, karena
keduanya SIRT1-dependent ( Araki et al., 2004 ) dan mekanisme -independent
dilaporkan ( Wang et al., 2005b ).

penyakit Alzheimer

Penyakit Alzheimer adalah penyakit neurodegenerative terminal, menyebabkan

kematian neuronal dan atrofi otak. Keunggulan patologis AD adalah kusut
intraseluler dan ekstraseluler plak di otak. Kusut, juga dikenal sebagai kusut
neurofibrillary, dibentuk oleh akumulasi protein tau larut, dan plak adalah
deposito dari -amyloid (A) peptida, biasanya terdiri dari 40-42 residu asam
amino.

Efek perlindungan dari SIRT1 terhadap AD awalnya diamati dalam studi CR, di
mana CR berkurang A dan generasi plak di otak tikus AD transgenik ( Patel et
al, 2005. ; Wang et al, 2005a. ). Demikian pula, pengurangan A juga melihat di
korteks monyet tupai berpuasa dan berbanding terbalik dengan tingkat SIRT1
( Qin et al., 2006a ). Studi ini menyiratkan bahwa SIRT1 terlibat dalam pelindung
saraf terhadap AD. Memang, studi terbaru menunjukkan bahwa aktivasi SIRT1
mengurangi kematian neuronal dan atrofi otak yang merupakan ciri khas dari AD
( Chen et al, 2005b. ; Qin et al, 2006b. ; Kim et al, 2007. ; Donmez et al,
2010. ; Min et al., 2010 ). Kekurangan SIRT1 dikaitkan dengan peningkatan
tingkat terfosforilasi-tau di neuron ( Min et al., 2010 ) dan jumlah kusut
neurofibrillary di otak AD ( Julien et al., 2009 ).

Selain itu, studi terbaru menunjukkan bahwa baik administrasi resveratrol atau
berlebih dari SIRT1 mengurangi tingkat A baik in vitro dan in vivo ( Chen et al,
2005b. ; . Qin et al, 2006b ; . Donmez et al, 2010 ). SIRT1 berlebih merangsang
produksi -secretase dalam neuron dan tikus oleh dua jalur: mengaktifkan
reseptor asam retinoat; (RAR Donmez et al, 2010. Dan menghambat rho-terkait,
melingkar-coil yang mengandung protein kinase 1 (ROCK1;) qin et al.,
2006b ). Peningkatan kadar -secretase meningkatkan proses normal Amyloid
prekursor protein (APP), yang mengarah ke generasi penurunan beracun
A. Selain itu, SIRT1 juga mengurangi jalur NF-kappaB di mikroglia dan
menurunkan tingkat A ( Chen et al., 2005b ). Secara bersama-sama, hasil ini
menetapkan bahwa SIRT1 melindungi terhadap AD oleh beberapa mekanisme,
termasuk penurunan tau dan tingkat mengurangi dari A.

penyakit Parkinson
Penyakit Parkinson adalah penyakit neurodegeneratif umum yang disebabkan
oleh kematian neuron dopaminergik dari substansia nigra di otak tengah. Gejala
utama dari PD adalah kekakuan, tremor, dan bradikinesia. Studi awal kami
menemukan bahwa CR atau penggunaan 2-deoxy- D -glucose, analog glukosa,
mengurangi hilangnya neuron dopaminergik pada tikus dan meningkatkan fungsi
motorik, menyiratkan bahwa SIRT1 mungkin terlibat dalam perlindungan ( Duan
dan Mattson, 1999 ) . Tingkat SIRT1 dalam neuron dopaminergik yang tajam
menurun pengobatan dengan neurotoksin, seperti rotenone, 6-
hydroxydopamine, -synuclein, atau 1-metil-4-fenil-1,2,3,6-tetrahydropyridine
(MPTP; Alvira et al, 2007. ; Pallas et al, 2008. ; Albani et al, 2009. ), yang adalah
agen banyak digunakan untuk model PD. Selain itu, SIRT1 berlebih ( Wareski et
al, 2009. Atau aktivasi dengan resveratrol () Okawara et al, 2007. ; Chao et al,
2008. ; . Albani et al, 2009 ) memperlambat kematian neuronal serta
neurodegeneration dalam model PD baik di vivo dan in vitro ( Donmez et al.,
2012 ), menunjukkan peran neuroprotektif SIRT1 terhadap PD. Tidak semua studi
menunjukkan peran protektif dari SIRT1, namun. Misalnya, tidak ada
perlindungan yang diamati pada model PD MPTP-diinduksi di SIRT1 tikus
transgenik ( Kakefuda et al., 2009 ). Namun demikian, meskipun kontroversi,
penelitian yang paling menunjukkan peran protektif dari SIRT1 terhadap PD,
meskipun mekanisme yang jelas.

Penyakit Huntington

Penyakit Huntington adalah autosomal penyakit keturunan yang dominan

dengan onset di usia menengah. Hal ini disebabkan oleh mutasi trinucleotide
ulangi pada gen huntingtin yang menghasilkan peningkatan jumlah residu
glutamin dalam N-terminus dari protein huntingtin yang menyebabkan agregasi
protein abnormal dan kematian akhirnya neuronal. Penelitian kami sebelumnya
menunjukkan bahwa CR bisa memperbaiki fenotip bermotor dan memperpanjang
kelangsungan hidup HD tikus ( Duan et al., 2003 ), menunjukkan bahwa jalur
yang berhubungan dengan metabolisme energi dapat mengubah perkembangan
penyakit pada penyakit. CR meningkatkan mitokondria biogenesis dengan
menginduksi endotel oksida nitrat sintase (eNOS), dan NO dapat mengaktifkan
gen SIRT1 ( Nisoli et al, 2005. ; Haigis dan Guarente 2006 ), yang merupakan
ortolog mamalia ragi Sir2, dan NAD sangat lestari + deacetylase protein
-tergantung. Selain itu, SIRT1 telah diusulkan untuk memediasi beberapa efek
menguntungkan dari CR ( Canto dan Auwerx, 2009 ; . Wakeling et al,
2009 ; Shimokawa dan Trindade, 2010 ; Chalkiadaki dan Guarente 2012 ).

Laporan pertama menunjukkan hubungan antara SIRT1 dan HD datang dari studi
oleh Parker et al. (2005) , yang menemukan bahwa berlebih dari Sir2.1 atau
pengobatan dengan resveratrol diselamatkan fenotipe disfungsi saraf yang
disebabkan oleh Polyglutamine mutan di Caenorhabditis elegans . Berbeda
dengan efek neuroprotektif Sir2.1 di C. elegans , Pallos et al. (2008) melaporkan
bahwa pengurangan 50% dari Sir2 diperpanjang kelangsungan hidup dan neuron
diawetkan mengandung fotoreseptor pada lalat mengekspresikan mutan
Htt. Menariknya, dalam sistem model terbang, berlebih dari Sir2 tidak
mengurangi mematikan atau tingkat degenerasi neuron yang disebabkan oleh
mutan Htt. Studi di kedua C. elegans dan Drosophila menunjukkan bahwa
hilangnya lengkap Sir2 adalah merusak di cacing ( Parker et al., 2005 ) dan
merusak dibandingkan dengan kerugian heterozigot di mutan lalat Htt-
menantang ( Pallos et al., 2008 ). Meskipun kehilangan heterozigot dari Sir2
adalah pelindung pada lalat, hilangnya heterozigot dari Sir2 di cacing tidak
diperiksa. Namun demikian, pengurangan Sir2 tidak mengubah rentang hidup
lalat tidak mengungkapkan Htt atau saudara mengekspresikan Htt. Beberapa
aspek dari peran sirtuins di umur di C. elegans dan Drosophila yang
kontroversial, dan penelitian telah menunjukkan bahwa Sir2 berlebih tidak
meningkatkan jangka hidup dan bahwa pembatasan diet meningkat umur lalat
secara independen dari dSir2 ( Burnett et al., 2011 ). Meskipun demikian,
berlebih dari Sir2 meningkatkan umur panjang lalat normal dan umur panjang
lalat yang sakit sedikit meningkat tinggi Sir2 ( Pallos et al., 2008 ). Hasil yang
berbeda mungkin disebabkan karena jumlah Sir2, status aktivasi, dan target hilir
yang berbeda yang terlibat. Hasil kontroversial menjamin penyelidikan lebih
lanjut dari peran SIRT1 dalam sistem mamalia.

Memang, dua studi independen oleh kelompok kami ( Jiang et al., 2011 ) dan
kelompok Krainc ini ( Jeong et al., 2011 ) menunjukkan bahwa modulasi tingkat
SIRT1 memiliki manfaat terapeutik dalam tiga model HD mouse yang berbeda,
dan target hilir diduga dari SIRT1 terlibat dalam hasil penyakit membaik juga
diidentifikasi. Kedua penelitian memberikan dukungan kuat untuk pandangan
bahwa SIRT1 memberikan efek menguntungkan pada model HD mouse, tetapi
juga menimbulkan pertanyaan penting. Hal ini dimungkinkan bahwa hasil yang
bertentangan tentang efek SIRT1 dalam model HD bisa dijelaskan dengan jalur
efektor yang berbeda atau mekanisme dan dengan efek tergantung pada
konteks atau tingkat yang berbeda dari aktivasi SIRT1. SIRT1 memiliki banyak
target, dan model yang berbeda dari HD menampilkan fenotipe yang berbeda
dengan mengaktifkan berbagai sasaran dan mekanisme. Oleh karena itu, tidak
mengherankan untuk mengamati data yang bertentangan, terutama pada
spesies yang berbeda dan model yang berbeda.

Amyotrophic Lateral Sclerosis

Amyotrophic lateral sclerosis adalah penyakit neurodegenerative kronis, fatal,

ditandai patologis dengan kematian neuron motorik di sumsum tulang belakang
dan korteks, mungkin disebabkan oleh kekurangan enzim superoxide dismutase
1 (defisiensi SOD-1; Rosen, 1993 ). Dalam model hewan ALS di mana bentuk
mutan dari defisiensi SOD-1 dinyatakan, tingkat SIRT1 yang diregulasi di motor
neuron ( Kim et al., 2007 ). SIRT1 berlebih melindungi neuron terhadap toksisitas
yang diinduksi oleh defisiensi SOD-1 mutan di kedua neuron berbudaya dan otak
tikus ( Kim et al., 2007 ). Perlindungan ini sesuai dengan peningkatan deasetilasi
p53. Resveratrol juga meningkatkan efek perlindungan dari SIRT1 dalam model
tikus ALS ( Kim et al, 2007. ; Markert et al, 2010. ), Tetapi beberapa dosis tampak
diperlukan untuk meningkatkan fungsi saraf dan meningkatkan umur panjang
tikus ( Markert et al., 2010 ).

multiple sclerosis
Multiple sclerosis adalah penyakit selubung myelin dengan lesi biasanya terletak
di otak, sumsum tulang belakang atau saraf kranial, dan yang paling umum, di
saraf optik. Penyebab multiple sclerosis tidak sepenuhnya diidentifikasi tapi
kemungkinan timbul dari etiologi autoimun; Oleh karena itu, secara tradisional
diperlakukan sebagai penyakit inflamasi. Baru-baru ini, bagaimanapun, multiple
sclerosis juga telah dianggap sebagai penyakit neurodegenerative karena co-
eksistensi kerusakan permanen aksonal, hilangnya neuron, dan kecacatan
neurologis pada pasien dengan penyakit ( Lassmann 2010 ; Shindler et al,
2010. ). Dalam model tikus dari multiple sclerosis, eksperimental
encephalomyelitis autoimun (EAE), SIRT1 aktivasi oleh SRT501 atau SRT1720
mempertahankan kepadatan aksonal, mencegah hilangnya neuron, dan
meningkatkan disfungsi neuronal ( Shindler et al., 2007 , 2010 ). SIRT1
penghambatan dengan Sirtinol melemahkan efek neuroprotektif SRT501
( Shindler et al., 2010 ), menunjukkan peran protektif dari SIRT1 dalam multiple
sclerosis. Namun, penelitian lebih lanjut diperlukan untuk sepenuhnya
menggambarkan peran SIRT1 dalam multiple sclerosis.

iskemia serebral

stroke iskemik adalah penyakit neurologis umum disebabkan oleh penurunan

tiba-tiba atau berhentinya aliran darah ke otak, menyebabkan infark. Manajemen
klinik stroke sulit dan obat-obatan saat ini harus diberikan dalam jendela waktu
yang terbatas setelah timbulnya stroke untuk memberikan manfaat
klinis. kandidat yang menjanjikan untuk strategi saraf termasuk penyesuaian
awal, hipotermia ringan, dan penggunaan senyawa kimia dan biologi
menargetkan mediator molekul penting kematian neuronal dan kelangsungan
hidup.

Efek neuroprotektif SIRT1 pertama kali dilaporkan pada pengkondisian iskemik

dan SIRT1 mengaktifkan senyawa resveratrol mengurangi cedera neuronal dari
hippocampus di iskemia serebral global dalam tikus. Peningkatan aktivitas SIRT1
juga terbukti menjadi mekanisme umum untuk efek protektif preconditioning dan
resveratrol ( Raval et al, 2006. ; Morris et al, 2011. ). Sirtinol, inhibitor aktivitas
SIRT1, menghapuskan pelindung saraf dari pengkondisian dan resveratrol ( Raval
et al., 2006 ), menunjukkan bahwa SIRT1 memainkan peran kunci dalam mediasi
pelindung saraf. Peran saraf ini kemudian didukung oleh dua studi terbaru
( Chong dan Maiese 2008 ; . Della-Morte et al, 2009 ) menunjukkan bahwa
aktivasi SIRT1 mengurangi cedera neuronal iskemik.

Studi lain menunjukkan bahwa, dalam budaya neuronal primer, pretreatment

dengan NAD + pretreatment nyata mengurangi kematian neuronal yang
disebabkan oleh kekurangan oksigen-glukosa, sebuah in vitro model iskemia
( Wang et al., 2008 ). SIRT1 diperlukan untuk NAD + pelindung saraf, seperti
NAD + pengobatan meregulasi ekspresi dan aktivitas SIRT1, dan SIRT1
knockdown melemahkan perlindungan dimediasi oleh NAD + ( Wang et al.,
2008 ). NAMPT berlebih mengurangi infark iskemik, sedangkan NAMPT
penghambatan memperburuk cedera iskemik. Efek perlindungan dari NAMPT
adalah SIRT1-tergantung, sebagai blok SIRT1 knockout perlindungan ( Wang et
al., 2011 ).

Meskipun bukti tersebut, kontroversi ada lebih dari efek perlindungan dari SIRT1
terhadap iskemia. Dalam sebuah penelitian dengan SIRT1 tikus transgenik, di
mana SIRT1 manusia diekspresikan di bawah kendali tikus neuron khusus
enolase promotor, tidak ada pelindung saraf diamati terhadap stroke SIRT1 dan
tipe liar tikus menunjukkan volume infark hampir tidak bisa dibedakan dan skor
defisiensi neurologis ( Kakefuda et al ., 2009 ). Perbedaan antara penelitian ini
dan yang lain mungkin karena tingkat tinggi berkelanjutan SIRT1, karena dapat
mengkonsumsi terlalu banyak atau bahkan menguras NAD + , yang bisa
membesar-besarkan cedera saraf ( Wang et al, 2008. ; . Kakefuda et al,
2009 ; Liu et al, 2009. ). Oleh karena itu, adalah mungkin bahwa
NAD + kekurangan dikompromikan efek neuroprotektif SIRT1. Dalam studi lain,
nicotinamide, senyawa yang menghambat aksi SIRT1, menunjukkan pelindung
saraf terhadap cedera iskemik, menyiratkan bahwa SIRT1 mungkin memiliki efek
yang merugikan terhadap stroke ( Chong et al., 2005 ). Namun, laporan ini
mungkin mengabaikan fungsi lain dari nicotinamide, termasuk dari prekursor
untuk NAD + sintesis. Bahkan, kelompok yang sama kemudian melaporkan
bahwa SIRT1 berlebih mencegah neuron dari apoptosis setelah stres oksidatif
( Chong dan Maiese 2008 ).

SIRT1 dalam Praktek Klinis

Senyawa SIRT1-mengaktifkan belum terbukti secara klinis berguna untuk

pengobatan penyakit neurodegenerative. Studi praklinis telah dilakukan dalam
berbagai model penyakit neurodegenerative, namun. Informasi yang diperoleh
dari studi tersebut bisa membuktikan menjadi berharga untuk merancang
molekul SIRT1-mengaktifkan yang mungkin lebih mungkin berguna secara
klinis. Penemuan molekul tersebut menjadi semakin penting, mengingat
keterbatasan manipulasi genetik dan kurangnya bukti tegas aktivasi SIRT1
tertentu dengan molekul prototipe seperti resveratrol ( Sauve, 2009 ).

Beberapa strategi yang rasional berdasarkan struktur protein yang tersedia dan
jalur katalitik telah dirancang untuk mengembangkan molekul kecil yang selektif
mengaktifkan sirtuins ( Sauve 2009 ). Salah satu strategi melibatkan merancang
molekul resveratrol-seperti, yang belum membuahkan hasil yang sukses
sebagai in vivo mekanisme aktivasi SIRT1 tidak sepenuhnya
dipahami. Pendekatan lain bertujuan untuk meningkatkan tingkat seluler dari
NAD + untuk mengaktifkan fungsi SIRT1. Pendekatan ini memiliki keuntungan
dari memanfaatkan jalur metabolisme alami untuk meningkatkan fungsi
SIRT1. Selain itu, terjadi secara alami metabolit menyajikan risiko paling
toksisitas. The khasiat dari agen yang telah digunakan untuk meningkatkan
NAD + masih dipertanyakan, namun, dan NAD +enhancement mempengaruhi
sejumlah jalur fisiologis lainnya, sehingga pendekatan ini tidak spesifik untuk
SIRT1. Strategi ketiga sedang dalam bukti-prinsip tahap ditunjuk nicotinamide
derepresi berdasarkan melawan efek penghambatan nicotinamide pada sirtuins
dengan merancang molekul yang bertentangan dengan
nikotinamida. Pendekatan ini masih dalam masa pertumbuhan dan belum
memberikan senyawa dengan potensi yang diinginkan, tetapi merupakan
strategi yang menarik untuk mengembangkan lebih lanjut. Rincian dari upaya
untuk menemukan molekul SIRT1-mengaktifkan baru-baru ini secara
komprehensif ditinjau oleh Blum et al. (2011) .

Perspektif masa depan SIRT1

Selama dekade terakhir, pemahaman kita tentang biologi sirtuins telah jauh
meningkat dari deskripsi aslinya sebagai NAD + -tergantung kelas III histone
deacetylase yang dapat mengontrol umur ragi. Yang menarik adalah penemuan
bahwa SIRT1 tidak hanya deacetylates histon, tetapi juga beberapa regulator
transkripsi terkenal, sehingga modulasi beragam proses biologis. Sebuah aspek
yang menarik adalah bahwa SIRT1 menengahi pelindung saraf terhadap kedua
penyakit neurologis akut dan kronis. Yang penting, aktivitas SIRT1 ditingkatkan
oleh senyawa-molekul kecil; Oleh karena itu, pengembangan aktivator-molekul
kecil dapat menyebabkan terapi baru terhadap penyakit saraf. Salah satu
mekanisme pelindung luas SIRT1 adalah untuk menekan genome-wide
transkripsi gen melalui histone deasetilasi. Selanjutnya, SIRT1 selektif menekan
gen yang terlibat dalam penyimpanan lemak, apoptosis, dan
peradangan. Menambah kompleksitas SIRT1-dimediasi kelangsungan hidup sel,
SIRT1 khusus mempromosikan transkripsi satu set gen yang berhubungan
dengan kelangsungan hidup sel, metabolisme energi, dan biogenesis
mitokondria. SIRT1 sehingga memiliki mekanisme multifaset dengan tujuan akhir
untuk meningkatkan kelangsungan hidup sel.

Meskipun dipelajari secara ekstensif, fungsi biologis SIRT1 tetap hanya sebagian
ditandai. Sehubungan dengan mekanisme kerja, ada beberapa faktor yang tidak
diketahui besar. Sebagai contoh, tidak diketahui bagaimana SIRT1 khusus
meningkatkan transkripsi gen menguntungkan sementara itu secara bersamaan
menekan transkripsi universal. Ini akan menarik untuk menentukan apakah
mekanisme yang sama ada untuk gen diregulasi oleh aktivasi SIRT1-dimediasi
transkripsi. Masalah lainnya adalah efek paradoks SIRT1. Misalnya, sedangkan
SIRT1 langsung menekan PPAR aktivitas transkripsi langsung ( Picard et al,
2004. , Itu juga mengaktifkan PGC-1) ( Nemoto et al, 2005. ; Rodgers et al,
2005. ), Yang dapat meningkatkan aktivitas transkripsional PPAR? ( Puigserver et
al., 1998 ).

Meskipun SIRT1 telah ditemukan saraf dalam berbagai studi, jelas dari
mekanisme patologis beragam diwujudkan dalam gangguan neurodegenerative
bahwa peran SIRT1 membutuhkan penelitian yang lebih rinci. Ketersediaan
struktur kristal dan analisis mekanistik rinci membantu dalam menemukan
modulator SIRT1, tapi akan menjadi nilai yang terbatas jika mereka gagal
mencapai uji klinis, sehingga menekankan pentingnya mengembangkan model
hewan yang kuat untuk menyelidiki mekanisme molekuler yang terlibat dalam
aktivasi SIRT1. Selain itu, efek negatif dari aktivasi SIRT1 dan penipisan energi
perlu penyelidikan lebih lanjut pada hewan model. Keberhasilan klinis senyawa
mengaktifkan sirtuin (STACs) dalam penyakit neurodegenerative bergantung
sangat pada pengembangan strategi baru dan merancang molekul berdasarkan
pada kimia sirtuin dan jalur molekuler diaktifkan oleh SIRT1. Sebuah studi baru-
baru ini sangat dilakukan oleh Hubbard et al. (2013) menunjukkan bahwa motif
hidrofobik tertentu dalam substrat SIRT1 seperti PGC-1 dan FOXO3a
memfasilitasi aktivasi SIRT1 oleh STACs, menyiratkan bahwa SIRT1 dapat
langsung diaktifkan melalui mekanisme alosterik umum untuk kimia beragam
STACs. Singkatnya, tidak ada keraguan bahwa SIRT1 memiliki potensi terapi yang
menjanjikan untuk gangguan neurodegenerative.

SIRT3, Energi Metabolisme dan Aging

Bagian ini merangkum studi tentang peran SIRT3 mitokondria dalam

metabolisme energi dan perlindungan terhadap stres oksidatif dan disfungsi
terkait usia (Gambar 2 ).

GAMBAR 2

GAMBAR 2. gambaran skematis dari target molekul SIRT3 dan peran

dalam mengatur metabolisme dalam mitokondria.

SIRT3 dan Mitokondria

Mitokondria adalah tidak hanya pembangkit tenaga listrik untuk produksi ATP
tetapi juga situs utama di mana spesies oksigen reaktif (ROS) yang dihasilkan
dan intrinsik apoptosis jalur pensinyalan dimulai ( Salminen et al.,
2008b ). Fungsi protein mitokondria yang diubah ketika mereka deasetilasi oleh
NAD + deacetylases mitokondria -tergantung, termasuk SIRT3, SIRT4, dan
SIRT5. Semua sirtuins mitokondria yang hadir dalam matriks mitokondria
( Howitz et al, 2003. ; Rogina dan Helfand, 2004 ). Sejak mitokondria
mengandung DNA mereka sendiri, faktor transkripsi, protein histon-seperti, dan
sistem sintesis protein, sirtuins mitokondria deacetylate satu set target dalam
mitokondria yang berbeda dari orang-orang dari protein nuklir ( Tissenbaum dan
Guarente, 2001 ; . Wood et al, 2004 ). Meskipun informasi mekanistik yang tepat
masih kurang, bukti yang muncul yang menunjukkan bahwa sirtuins mitokondria
melindungi terhadap stres oksidatif ( Braunstein et al., 1993 ; . Baur et al, 2006 ).

Di antara sirtuins mitokondria, fungsi SIRT3 telah dicirikan detail terbesar. Studi
awal tikus SIRT3-kekurangan menunjukkan bahwa hilangnya SIRT3, tapi tidak
SIRT4 atau SIRT5, menyebabkan hyperacetylation protein dramatis dalam
mitokondria, menunjukkan SIRT3 yang aktivitas deacetylase mitokondria utama
( Dali-Youcef et al., 2007 ). Pada manusia, full-length SIRT3 adalah protein 44-kD
dengan N-terminal urutan penargetan mitokondria yang merupakan enzimatis
aktif in vitro . Hal ini proteolitik diproses dalam mitokondria untuk dewasa 28 kD
katalitik deacetylase aktif ( Onyango et al, 2002. ; Schwer et al., 2002 ). Mouse
pertama SIRT3 cDNA urut diidentifikasi dikodekan protein 28-kD kurang N-
terminal mitokondria urutan penargetan ( Yang et al., 2000 ). Beberapa
penelitian terbaru telah mengidentifikasi isoform lagi dari SIRT3 murine
pengkodean protein 37 kD, bagaimanapun, yang dapat diimpor ke mitokondria
dan diolah menjadi matang 28 protein kD ( Cooper et al, 2009. ; Jin et al,
2009. ; Bao et al, 2010. ; . Yang et al, 2010 ). Apakah fraksi aktif SIRT3 ada
mitokondria luar dan memodifikasi protein ekstra-mitokondria masih
kontroversial.

SIRT3 dan Metabolik Homeostasis

Data yang muncul menunjukkan bahwa salah satu fungsi utama dari SIRT3
adalah regulasi aktivitas rantai transpor elektron mitokondria untuk
mempertahankan homeostasis energi. Sumber energi utama di mitokondria
adalah piruvat, produk glikolisis. Atau, mitokondria juga membakar asam lemak,
asam amino, dan asetat ketika piruvat kekurangan. Untuk katabolisme asam
lemak, rantai panjang asil koenzim A dehidrogenase (LCAD) adalah enzim kunci
yang memecah asam lemak dan menghasilkan asetil-CoA, merangsang -
oksidasi. Dalam SIRT3 tikus knockout, LCAD adalah hyperacetylated di Lys42,
mengarah ke penurunan aktivitas enzim, -oksidasi, dan tingkat ATP ( Hirschey
et al., 2010 ). Menariknya, tikus-tikus ini tidak mentolerir paparan dingin selama
puasa ( Hirschey et al., 2010 ). SIRT3 langsung deacetylates LACD di Lys42 dan
meningkatkan aktivitas LACD ( Hirschey et al., 2010 ). Selain itu, SIRT3 dapat
mempromosikan -oksidasi melalui beberapa mekanisme, seperti dengan
deacetylating enzim -oksidasi lainnya, termasuk rantai pendek L -3-
hydroxyacyl-CoA dehidrogenase dan koenzim sangat panjang rantai asil A
dehidrogenase ( Hallows et al., 2011 ), memfasilitasi adaptasi mitokondria untuk
bahan bakar perubahan.

Glukosa adalah sumber energi utama bagi sel-sel. Ketika ketersediaannya

terbatas, bagaimanapun, bahan bakar alternatif menjadi semakin penting bagi
kelangsungan hidup sel. Langkah pertama glikolisis adalah konversi glukosa
menjadi glukosa-6-fosfat, reaksi dikatalisis oleh hexokinases. Hal ini melaporkan
bahwa SIRT3 deacetylates cyclophilin D ( Hafner et al, 2010. ; Shulga et al,
2010. ), Yang mengarah ke pemisahan heksokinase II dan mitokondria,
menurunkan metabolisme glukosa, dan menstimulasi fosforilasi oksidatif
( Shulga et al, 2010. ).

Asetat berasal dari asam asetat dan alkohol juga digunakan sebagai bahan bakar
mitokondria, meskipun ini hanya terjadi dalam keadaan ekstrim deplesi
nutrisi. Pada langkah asetat awal diubah menjadi asetil-CoA dikatalisis oleh
sintetase asetil-CoA. Asetil-CoA sintetase 2 adalah bentuk mitokondria
enzim. SIRT3 deacetylates asetil-CoA sintetase 2 dan meningkatkan aktivitasnya,
yang mengarah ke peningkatan produksi asetil-CoA ( Hallows et al,
2006. ; Schwer et al, 2006. ). Asetil-CoA dari asam lemak dan asetat serta -
ketoglutarat dari asam amino dapat memasuki siklus Krebs. Kedua reaksi
ditingkatkan oleh SIRT3 ( Hallows et al, 2006. ; . Schwer et al, 2006 ; . Lombard
et al, 2007 ; . Schlicker et al, 2008 ). Selain itu, SIRT3 langsung merangsang
siklus Krebs. Langkah ketiga dari siklus adalah konversi isositrat 6-karbon untuk
5-karbon -ketoglutarat, proses dikatalisasi oleh isositrat dehidrogenase 2
(IDH2). Sebuah penelitian baru menunjukkan bahwa SIRT3 langsung
deacetylates dehidrogenase ini untuk meningkatkan aktivitasnya ( Someya et al.,
2010 ).

NADH dehidrogenase 1 alpha subcomplex subunit 9 (NDUFA9) adalah enzim

kompleks mitokondria saya yang asetat di Lys370 ( Kim et al., 2006 ). SIRT3 fisik
berinteraksi dengan NDUFA9 dan deacetylates itu. SIRT3 KO meningkatkan
asetilasi dan mengurangi aktivitas kompleks I ( Ahn et al., 2008 ), menunjukkan
bahwa SIRT3 adalah pengatur positif dari kompleks I. Complex II, juga dikenal
sebagai suksinat dehidrogenase, terdiri dari empat subunit, termasuk
flavoprotein yang suksinat dehidrogenase subunit A (SdhA). Dalam SIRT3 tikus
knockout, SdhA adalah hyperacetylated di beberapa residu lisin, dan
menunjukkan penurunan aktivitas kompleks II ( Cimen et al., 2010 ). Berlebih
SIRT3 membalikkan asetilasi SdhA dan meningkatkan kompleks aktivitas II
( Cimen et al., 2010 ), menunjukkan bahwa SdhA adalah substrat SIRT3, dan
bahwa SIRT3 juga regulator positif dari kompleks II.

SIRT3 juga dilaporkan untuk mengikat alpha subunit 1 dari partikel F1 ATP
synthase ( UU et al., 2009 ), namun fungsi ini tidak jelas. Secara bersama-sama,
hasil ini menunjukkan bahwa SIRT3 mempromosikan generasi ATP melalui
peningkatan aksi dari beberapa enzim yang terlibat dalam metabolisme
energi. Selanjutnya mendukung peran ini, tikus SIRT3-KO menunjukkan asetilasi
besar protein mitokondria, dan telah mengurangi tingkat ATP pada awal dan
selama stres seluler ( Ahn et al., 2008 ).

Mitokondria juga situs utama untuk generasi ROS, superoksida, dan juga di mana
superoksida yang dismuted oleh mitokondria MnSOD. Laporan terbaru
menunjukkan bahwa SIRT3 deacetylates MnSOD di Lys122 dan meningkatkan
aktivitasnya, mengurangi stres oksidatif dan radiasi pada tikus ( Qiu et al,
2010. ; . Tao et al, 2010 ). Berlebih dari SIRT3 melindungi HEK293 dari stres
oksidatif dan mencegah kematian sel koklea yang berkaitan dengan usia pada
tikus ( Someya et al., 2010 ). Secara keseluruhan, ini menunjukkan anti-oksidatif
dan peran neuroprotektif SIRT3. Data baru menunjukkan bahwa SIRT3
deacetylase memainkan peran kunci dalam memperkuat mitokondria pertahanan
anti-oksidan selama CR ( Qiu et al, 2010. ; . Someya et al, 2010 ).

Dalam studi masa depan, itu akan menarik untuk menentukan mekanisme
dimana asetilasi elektron subunit rantai transpor mempengaruhi generasi
ATP. Hal ini juga penting untuk menjelaskan mengapa hal ini mungkin diinginkan
di bawah beberapa kondisi fisiologis untuk downregulate elektron aktivitas rantai
transportasi melalui peningkatan asetilasi. Sebagai kerut menambahkan, SIRT3
negatif mengatur terjemahan dalam mitokondria oleh deacetylating yang
MRPL10 protein ribosom, fungsi yang diusulkan untuk mengurangi respirasi
( Yang et al., 2010 ).
Tingkat SIRT3 meningkat dalam jaringan adiposa, otot rangka, dan hati selama
CR ( Shi et al, 2005. ; Palacios et al, 2009. ; Schwer et al, 2009. ; Hirschey et al,
2010. ), Dan sebaliknya penurunan respon untuk tinggi lemak makan ( Palacios
et al, 2009. ; . Bao et al, 2010 ; . Kendrick et al, 2011 ). Data ekspresi ini
menunjukkan bahwa SIRT3 mungkin memainkan peran dalam penanggulangan
homeostasis metabolik. Pada mamalia, dua studi independen menunjukkan
bahwa SIRT3 berinteraksi dengan dan deacetylates asetil-CoA sintetase 2
(AceCS2) di situs aktif lisin untuk mempromosikan aktivitas AceCS2 ( Hallows et
al, 2006. ; . Schwer et al, 2006 ). Dalam kondisi makan, mayoritas asetil-CoA
dihasilkan melalui metabolisme piruvat oleh piruvat dehidrogenase kompleks
(PDC) dan asam lemak -oksidasi, sebagian besar melewati kebutuhan untuk
AceCS2. Dalam hal ini, penelitian tikus AceCS2-kekurangan mengungkapkan
bahwa AceCS2 secara khusus diperlukan untuk homeostasis metabolik ketika
tikus diberi makan karbohidrat / lemak tinggi rendah (LC / HFD); Hewan AceCS2-
kekurangan pada dasarnya normal pada diet chow tapi menunjukkan kenaikan
miskin berat badan, hipotermia, hipoglikemia, dan gangguan kelangsungan
hidup pada LC / HFD ( Sakakibara et al., 2009 ). Agaknya peran SIRT3 dalam
mengatur AceCS2 juga bisa menjadi penting selama berpuasa, ketika asetat
dapat digunakan sebagai sumber energi dalam jaringan ekstrahepatik ( Hirschey
et al., 2010 ). Memang, SIRT3 baru-baru ini telah ditunjukkan untuk deacetylate
dan mengaktifkan 3-hydroxy-3-methylglutaryl-CoA sintase 2 (HMGCS2), enzim
mitokondria yang mengubah asetil-CoA menjadi badan keton (asetoasetat, -
hidroksibutirat, dan aseton) dalam hati dalam kondisi puasa, yang pada
gilirannya dapat digunakan sebagai sumber energi dalam jaringan tertentu
termasuk otak ( Shimazu et al., 2010 ). Tikus SIRT3-kekurangan tidak dapat
menghasilkan tingkat normal badan keton pada puasa.

SIRT, umur dan Age-Associated Fenotipe

Peran SIRT3 dalam penuaan adalah cukup menarik karena tampaknya untuk
menekan ROS, salah satu penyebab berkontribusi terhadap proses
penuaan. Selain elucidating peran dalam mengatur jalur biokimia tertentu dalam
mitokondria, ada minat yang besar dalam pengujian apakah SIRT3 mungkin
memodulasi fenotipe terkait usia, atau memang umur pakai sendiri. Dalam hal
ini, beberapa penelitian telah menghubungkan polimorfisme dalam SIRT3 lokus
genom untuk panjang umur manusia, meskipun orang lain telah gagal untuk
menunjukkan hubungan ini ( Rose et al, 2003. ; Bellizzi et al, 2005. , 2007 ; .
Lescai et al, 2009 ) . Sebuah polimorfisme terkait dengan penurunan ekspresi
SIRT3 mRNA hadir di kohort laki-laki muda tapi tidak lama, menunjukkan bahwa
mengurangi ekspresi SIRT3 dapat merugikan kelangsungan hidup di usia tua
( Bellizzi et al., 2005 , 2009 ). Pada individu menetap, ekspresi protein SIRT3
menurun dengan usia di mitokondria otot rangka, bersamaan dengan penurunan
fungsi pernafasan ( Lanza et al., 2008 ).

Terkait usia gangguan pendengaran (ARHL) adalah masalah umum pada orang
tua, terjadi akibat kehilangan sel dan perubahan degeneratif lainnya di
koklea. Sebuah studi yang elegan telah mapan peran SIRT3 dalam mencegah
ARHL ( Someya et al., 2010 ). Salah satu mekanisme yang SIRT3 memediasi efek
ini adalah dengan deasetilasi IDH2 ( Schlicker et al, 2008. ; . Someya et al,
2010 ), yang mengubah isositrat ke alfa-ketoglutarat bersamaan dengan
pengurangan NADP + . NADPH pada gilirannya memungkinkan regenerasi
berkurang glutathione untuk mempromosikan pertahanan oksidatif
mitokondria. Menanggapi CR, tikus wild type, namun hewan tidak SIRT3-
kekurangan, menunjukkan peningkatan kadar NADPH, meningkat glutation
tereduksi dalam mitokondria, dan penurunan kerusakan DNA dalam koklea dan
jaringan lain. Dalam sel kultur jaringan, overekspresi SIRT3 atau IDH2 melindungi
terhadap stres akibat kematian sel oksidatif, dan dua protein bersama-sama
memiliki efek pro-hidup sinergis. Hasil ini tidak mengesampingkan kemungkinan
bahwa SIRT3 dapat memodifikasi substrat lain selain IDH2 untuk mencegah AHRL
selama CR. Demikian pula, Qiu et al. (2010) melaporkan bahwa tikus SIRT3-
kekurangan gagal untuk menekan kadar ROS dan kerusakan makromolekul
selama CR. Mereka menemukan bahwa SIRT3 langsung deacetylates SOD2 untuk
meningkatkan aktivitas selama CR, sedangkan tikus SIRT3-kekurangan tidak
menunjukkan SOD2 deasetilasi dalam menanggapi diet ini ( Qiu et al.,
2010 ). Secara keseluruhan, makalah ini menunjukkan peran penting untuk SIRT3
dalam menekan kerusakan oksidatif dan sekuel negatif selama CR. Ini masih
harus dilihat bagaimana SIRT3, atau sirtuins mitokondria lainnya, mungkin
mempengaruhi fenotip lainnya dari penuaan dan / atau efek dari CR. Penurunan
insulin serum dan trigliserida yang biasanya terjadi selama CR tidak diamati
pada tikus SIRT3-kekurangan ( Someya et al., 2010 ), yang menyiratkan bahwa
SIRT3 memainkan tambahan, peran uncharacterized di adaptasi untuk rejimen
diet ini. Sebuah studi terbaru menunjukkan bahwa peningkatan regulasi SIRT3
memang membalikkan penuaan terkait degenerasi sel induk hematopoietik
( Brown et al., 2013 ), dan SIRT3 dapat mempromosikan organisme umur
panjang dengan mempertahankan integritas sel induk spesifik jaringan.

SIRT3 dan mitokondria Protein Asetilasi: Pertanyaan yang belum

terselesaikan

Asetilasi protein mitokondria memainkan peran utama dalam mengatur fungsi

organel ini. Meskipun kemajuan pesat di daerah ini, masih banyak pertanyaan
yang beredar tetap yang tidak diragukan lagi akan memberikan jalan bermanfaat
untuk penelitian selama bertahun-tahun yang akan datang. Secara khusus,
bagaimana protein mitokondria asetat di tempat pertama saat ini tidak
diketahui. Identitas acetyltransferases mitokondria diduga masih sulit
dipahami; identifikasi protein tersebut akan merupakan langkah maju yang besar
dalam bidang ini. Atau, atau di samping asetilasi enzimatik dalam mitokondria,
protein mitokondria pada prinsipnya bisa asetat luar organel ini, sebelum atau
bersamaan dengan impor mitokondria atau bahkan asetat non-enzimatik. Model-
model yang terakhir tidak akan mengizinkan siklus cepat asetilasi / deasetilasi
protein mitokondria untuk mengatur fungsi protein target dalam menanggapi
tantangan lingkungan bervariasi. Sebaliknya, setelah deasetilasi, pemulihan
status asetilasi akan membutuhkan sintesis protein baru. Model tersebut dapat
dibedakan melalui eksperimen pulsa-mengejar menilai asetilasi protein
mitokondria baru disintesis sebelum dan sesudah impor mitokondria.
Demikian pula, bagaimana kegiatan SIRT3 diatur dalam mitokondria yang tidak
sepenuhnya dipahami. SIRT3 membutuhkan NAD + , dan karena itu mitokondria
NAD + tingkat memainkan peran penting dalam mengatur fungsi SIRT3
mitokondria. Peningkatan generasi NADH dari NAD + yang terjadi dengan HFD
dan menyebabkan fungsi SIRT3 berkurang mungkin menjelaskan peningkatan
asetilasi protein mitokondria global diamati selama diet ini, karena bisa
meningkatkan kadar asetil-CoA, substrat untuk acetyltransferases ( Kendrick et
al., 2011 ) . Ini meningkat asetilasi keseluruhan dapat mewakili efek bersih dari
peningkatan aktivitas acetyltransferase ditumpangkan pada fungsi SIRT3
tinggi; alternatif, beberapa spesies protein hyperacetylated selama CR atau
kondisi lain mungkin tidak substrat untuk SIRT3 mitokondria. Aktivitas SIRT3 dan
sirtuins mitokondria lainnya mungkin dipengaruhi oleh kondisi lain selain
NAD +tingkat, seperti modifikasi pasca-translasi atau interaksi dengan protein
regulator.

Saat ini belum jelas apakah intervensi yang berdampak pada asetilasi banyak
protein mitokondria - kekurangan SIRT3, seperti CR, HFD - menyebabkan
modifikasi set umum dari protein di situs lisin yang sama, atau apakah
tanggapan ini disesuaikan dengan gangguan lingkungan yang
berbeda . Demikian pula, masih belum jelas apakah sirtuins mitokondria berbagi
target dan / atau fungsi umum yang sama. Pertanyaan ini bisa diatasi pada tikus
atau sel dengan kekurangan sirtuin mitokondria atau knockdowns. Mengingat
bahwa SIRT3 deacetylates banyak protein di mitokondria serta menekan
beberapa fenotipe terkait usia, itu akan menarik untuk menguji apakah asetilasi
perubahan protein mitokondria dengan usia, baik secara individual maupun
secara global, dan apakah pencegahan efek ini mungkin memiliki efek yang
menguntungkan pada rentang kesehatan atau bahkan umur.

Selain itu, sedangkan dampak fungsional asetilasi pada beberapa target protein
individu jelas, pemahaman global tentang bagaimana SIRT3 mempengaruhi
aktivitas jalur metabolisme dalam mitokondria, sel, jaringan, dan organisme
secara keseluruhan masih kurang. Jawaban untuk ini dan terkait pertanyaan
yang melibatkan SIRT3 dan asetilasi protein mitokondria tidak diragukan lagi
akan mengungkapkan aspek-aspek novel biologi mitokondria, dan mungkin
akhirnya memberikan dasar untuk strategi terapi baru untuk berbagai gangguan.

Perspektif lanjut

Dalam dekade terakhir, fungsi sirtuins mamalia telah diteliti secara lebih rinci
daripada sebelumnya, dan kami sekarang memiliki pemahaman molekul yang
jauh lebih baik dari beberapa peran keluarga yang unik ini enzim bermain di
tampaknya setiap proses biologis. Ada sedikit keraguan bahwa sirtuins telah
muncul sebagai modulator kritis tanggapan adaptif metabolik, dan kegiatan
mereka telah dikaitkan dengan kelainan metabolisme serta neurodegeneration
terkait usia. Namun, pertanyaan kunci akan tetap peneliti sibuk di tahun-tahun
mendatang. Kami masih memiliki pemahaman yang buruk tentang mekanisme
molekuler yang mengatur ekspresi dan aktivitas sirtuins, dan rangsangan yang
tepat yang mengatur protein ini, dan apakah kegiatan sirtuins yang berbeda
diatur secara terkoordinasi. Dengan kata lain, apakah ada cross-talk antara
sirtuins? Akan masa depan studi semen argumen bahwa sirtuins, memang,
modulator kritis umur? Ulasan ini telah difokuskan pada SIRT1 dan SIRT3. Bahkan
sedikit yang diketahui tentang sirtuins lainnya. Penyelidikan lebih lanjut ke dalam
sasaran dan fungsi keluarga ini unik sirtuins akan membantu mengembangkan
strategi baru untuk perlindungan terhadap dan pemulihan dari penyakit
neurologis umum yang berhubungan dengan penuaan dan meningkatkan
penuaan sukses.

Benturan Pernyataan Tujuan

penulis menyatakan bahwa penelitian ini dilakukan dalam tidak adanya

hubungan komersial atau keuangan yang dapat ditafsirkan sebagai potensi
konflik kepentingan.

REVIEW ARTICLE

Front. Aging Neurosci., 23 July 2013 | https://doi.org/10.3389/fnagi.2013.00036

Sirtuins: from metabolic regulation to brain aging

Wenzhen Duan1,2,3*
1
Division of Neurobiology, Department of Psychiatry and Behavioral
Sciences, Johns Hopkins University School of Medicine, Baltimore, MD, USA
2
Department of Neuroscience, Johns Hopkins University School of
Medicine, Baltimore, MD, USA
3
Program in Cellular and Molecular Medicine, Johns Hopkins University
School of Medicine, Baltimore, MD, USA

Brain aging is characterized by progressive loss of neurophysiological functions

that is often accompanied by age-associated neurodegeneration. Calorie
restriction has been linked to extension of lifespan and reduction of the risk of
neurodegenerative diseases in experimental model systems. Several signaling
pathways have been indicated to underlie the beneficial effects of calorie
restriction, among which the sirtuin family has been suggested to play a central
role. In mammals, it has been established that sirtuins regulate physiological
responses to metabolism and stress, two key factors that affect the process of
aging. Sirtuins represent a promising new class of conserved deacetylases that
play an important role in regulating metabolism and aging. This review focuses
on current understanding of the relation between metabolic pathways involving
sirtuins and the brain aging process, with focus on SIRT1 and SIRT3. Identification
of therapeutic agents capable of modulating the expression and/or activity of
sirtuins is expected to provide promising strategies for ameliorating
neurodegeneration. Future investigations regarding the concerted interplay of
the different sirtuins will help us understand more about the aging process, and
potentially lead to the development of therapeutic approaches for the treatment
of age-related neurodegenerative diseases and promotion of successful aging.
Introduction

The brain, similar to other organs, undergoes a gradual decline in energy

metabolism during aging (Drew and Leeuwenburgh, 2004; Navarro and Boveris,
2007; Boveris and Navarro, 2008; Swerdlow, 2011). Since neurons require large
amounts of energy for the firing of action potential, neurotransmission, and other
processes, the age-related decline in metabolism contributes to the cognitive
declines associated with aging (Biessels and Kappelle, 2005; Boveris and
Navarro, 2008). Aging is also a risk factor for age-associated diseases such as
neurodegenerative disorders. These diseases may occur when neurons fail to
respond adaptively to an age-related decline in basal metabolic rates and in
energy-driven tasks, such as neuromuscular coordination, cognitive performance,
and environmental awareness (Swerdlow, 2007). In the past decade, the function
of mammalian sirtuins, evolutionarily conserved nicotinamide adenine
dinucleotide (NAD)-dependent protein deacetylases/ADP-ribosyltransferases, has
been investigated in greater detail, and we now have a much better molecular
understanding of the multiple roles that this unique family of enzymes plays in
aging and seemingly every biological process. There is little doubt that sirtuins
have emerged as critical modulators of metabolic adaptive responses, and their
activities have been linked to multiple diseases, from metabolic abnormalities to
neurodegeneration.

Sirtuins were originally identified as one of the genes that regulate the mating
types of budding yeast, Saccharomyces cerevisiae, and named silent information
regulator 2 (Sir2) in lower organisms (Klar and Fogel, 1979). Following the first
publication describing a role for yeast Sir2 in promoting longevity (Kaeberlein et
al., 1999), many investigations focused on elucidating whether sirtuins might
play similar roles in other organisms. Sirtuins have been shown to regulate
lifespan in lower organisms, including yeast, nematodes, and fruit flies (Haigis
and Guarente, 2006), although their role in worm and fly lifespan has recently
been debated (Burnett et al., 2011; Viswanathan and Guarente, 2011). Most of
these studies have described a key role for SIRT1 in regulating the metabolic
response to calorie restriction (CR; Canto and Auwerx, 2009), a dietary
intervention that robustly extends life span across numerous species. However,
whole body overexpression of SIRT1 in mice does not affect life span (Herranz et
al., 2010). Nevertheless, SIRT1 does appear to promote healthy aging by
protecting against several age-related pathologies, such as facilitating insulin
sensitivity, elevating glucose production, reducing oxidative stress, potentiating
activity of brain-derived neurotrophic factor (BDNF) transcriptional factor cAMP
response element-binding protein (CREB; Guarente and Franklin, 2011).

Mammals have seven sirtuins (SIRT17) which are found in different subcellular
locations, including the nucleus (SIRT1, SIRT6, and SIRT7), cytosol (SIRT2), and
mitochondria (SIRT3, SIRT4, and SIRT5). Most of the studies have described a key
role for SIRT1 in regulating the metabolic response to CR (Canto and Auwerx,
2009), a dietary regimen involving reduced 3040% calorie intake compared to
normal calorie intake, that resulted in extended lifespan and reduced
development of morbidity with aging (Jiang et al., 2000; Masoro, 2000; Sinclair,
2002; Koubova and Guarente, 2003). Calorie restriction is the only intervention
that has consistently been shown to delay the onset, slow the progression of
age-related disease, and extend lifespan in short-lived species, as well as in long-
lived non-human primates, suggesting that similar mechanisms would be
operative in humans. Whole body overexpression of Sirt1 in mice does not affect
lifespan (Herranz et al., 2010). Nevertheless, SIRT1 promotes healthy aging by
preventing age-associated pathologies (Guarente and Franklin, 2011). Another
strong link between mammalian sirtuins and the anti-aging effects of CR was
provided by SIRT3, which mediates the prevention of age-related hearing loss
(Someya et al., 2010). SIRT3 is required for the CR-mediated reduction of
oxidative damage in multiple tissues via regulation of the glutathione antioxidant
system (Someya et al., 2010).

In this review, we focus on the effects of SIRT1 and SIRT3 on metabolic regulation
and their anti-aging activity in brain, and further discuss potential
pharmacological approaches to remedy and prevent age-associated neurological
disorders by targeting sirtuins.

SIRT1, Metabolism and Brain Aging

Distribution of SIRT1 in the Brain

During mouse embryogenesis, SIRT1 is highly expressed in the brain, spinal cord,
and dorsal root ganglion, with the peak expression at E4.5 (Salminen and
Kaarniranta, 2012). SIRT1 is also expressed in the adult brain, with high levels in
the cortex, hippocampus, cerebellum, and hypothalamus, and low levels in white
matter (Singh, 2004). Among the various cell types of brain, SIRT1 is
predominantly, if not exclusively, expressed in neurons (Singh, 2004; Adler et al.,
2007; Salminen and Kaarniranta, 2012). The only exception is that SIRT1 is found
in microglia when co-cultured with neurons (Schmitz et al., 2004). At the
subcellular level, SIRT1 is viewed as a nuclear protein (Chen and Greene, 2003).
Yet it is reported that SIRT1 has both nuclear import and export sequences, and
that SIRT1 is present in the cytosolic fraction of mouse brain, although its
cytosolic function is just beginning to be elucidated (Chen et al., 2005b; Lee et
al., 2008; Hardie, 2011).

SIRT1 Mediates Metabolic Benefits Under CR

SIRT1 contains 747 amino acids in humans, with a predicted molecular weight of
81 kDa and a measured one of 120 kDa. In addition to histones, SIRT1 also
deacetylates a number of non-histone substrates, including p53 (Luo et al.,
2001)and peroxisome proliferator-activated receptor (PPAR) coactivator-1
(PGC-1; Nemoto et al., 2005), and FOXO (Xiong et al., 2011), nuclear factor -
light-chain-enhancer of activated B cells (NF-B; Salminen et al., 2008b). SIRT1 is
drawing even more attention since it is considered to be one of the determining
factors in lifespan extension induced by CR, a phenomenon observed in
phylogenetically diverse organisms including yeast, worm, fruit fly, and mouse
(Kaeberlein et al., 1999; Tissenbaum and Guarente, 2001; Howitz et al.,
2003; Rogina and Helfand, 2004). Its beneficial roles are further supported by the
findings that putative SIRT1-activating compounds, such as resveratrol, also
promote longevity in several species, including yeast (Howitz et al., 2003), worm
(Wood et al., 2004), and mouse (Baur et al., 2006), making it an anti-aging
target.

The effects of SIRT1 on longevity rely on its enzymatic activity of deacetylation of

histone and non-histone substrates. While the deacetylation of histones leads to
their interaction with DNA and consequent gene silencing (Braunstein et al.,
1993; Sauve et al., 2006; Dali-Youcef et al., 2007), the deacetylation of non-
histone proteins has a wide range of biological effects, including metabolic
adjustment, survival promotion, and autophagy (Campisi, 2005; Dali-Youcef et
al., 2007; Brooks and Gu, 2009; Madeo et al., 2010). For example, SIRT1 inhibits
p53 (Luo et al., 2001), reducing its pro-apoptotic effect. It also inhibits NF-B
(Yeung et al., 2004), reducing its pro-inflammatory effects. In contrast, SIRT1
activates a transcriptional coactivator, PGC-1 (Nemoto et al., 2005), leading to
increased glucose levels, insulin sensitivity, and mitochondrial biogenesis.
Together, these and other effects, contribute to the longevity evoked by CR
(Figure 1).

FIGURE 1

FIGURE 1. Schematic diagram of anti-aging mechanism activated by

SIRT1 and calorie restriction. Calorie restriction upregulates the activity and
levels of SIRT1, SIRT1 deacetylates its substrates, including histone and non-
histone molecules, and improve genome stability, prevent apoptosis and
inflammation, and increases mitochondrial biogenesis, insulin sensitivity and
glucose tolerance. All these effects result in longevity.

These metabolic changes and the cytoprotective involvement of CR are generally

considered to occur in non-neural organs, such as the liver, pancreas, muscle,
and fat tissues (Brooks and Gu, 2009; Imai and Guarente, 2010). However,
recent studies suggest that the hypothalamus may also contribute to the
longevity effects of SIRT1 and CR via coordination of neurobehavioral and
neuroendocrine changes, including body temperature, appetite, and overall
physical activity (Dietrich et al., 2010; Satoh et al., 2010). SIRT1 is abundantly
expressed in several regions in the hypothalamus of mice, especially in the
arcuate, paraventricular, ventro- and dorsomedial nuclei; and CR increases SIRT1
levels in the hypothalamus, which increases body temperature, food intake, and
physical activity (Ramadori et al., 2008; Dietrich et al., 2010; Satoh et al., 2010).
SIRT1 appears to be required for the aforementioned behavioral changes, which
are prevented if SIRT1 is knocked out or inhibited (Chen et al., 2005a; Satoh et
al., 2010). In addition to the hypothalamus, SIRT1 is also expressed in other
regions of the brain, including the cortex, striatum, and hippocampus (Ramadori
et al., 2008). Shortly after this finding, a neuroprotective role of SIRT1, has been
reported (Tang, 2009; Morris et al., 2011).

SIRT1 and Factors Involved in CR and Aging

SIRT1 has been found to delay aging and promote longevity by regulating the
activity of key cellular proteins like p53, FOXO and Ku70 that are involved in
either apoptotic processes or cellular repair mechanisms. SIRT1 may thus
promote health and longevity partly by either decelerating cell death and/or by
boosting repair mechanisms in the cells (Wang et al., 2010).

It has become increasingly evident that the salutary effects of the CR, are in part
due to the promotion of sirtuins (Wang et al., 2010). The expression levels of
SIRT1 increase upon CR in several rodent and human tissues, including white
adipose, liver, skeletal muscle, brain, and kidney. Levels of NAD have been
shown to rise in liver cells under CR-like conditions, which in turn induces
expression of SIRT1 (Rodgers et al., 2005). SIRT1 ends up consuming NAD+ as a
result of its deacetylase activity, generating nicotinamide, an inhibitor of its own
activity. NAD+ is known to protect neurons (Liu et al., 2009) and thus by
increasing the levels of NAD+, CR may preserveSIRT1 activity. SIRT1 also
activates PGC1 (Rodgers et al., 2005) which results in mitochondrial biogenesis
(Liu et al., 2009). A decline in mitochondrial activity is thought to be causative in
many age-related diseases (Petersen et al., 2003; Singh, 2004). CR evokes
improvements in mitochondrial activity similar to those of SIRT1. Therefore, it is
possible that small-molecule modulators of SIRT1 may act on the same pathways
as those modified by CR, and thus have potential to mitigate age-related
diseases (Lavu et al., 2008).

SIRT1 interacts with and modulates other key factors involved in mammalian
aging, such as NF-B that controls a low-grade systemic inflammation along with
human aging process-inflamm-aging (Salminen et al., 2008a), mammalian target
of rapamycin (mTOR; Finley and Haigis, 2009), AMP-activated protein kinase
(AMPK; Salminen and Kaarniranta, 2012), therefore controls the gin process. The
aging process involves changes in immune regulation; NF-B signaling is the
master regulator of the immune system. Inhibition of NF-B signaling in aged
mice reverted the tissue characteristics and global gene expression to those of
young mice (Adler et al., 2007). The function of the NFB complex can be
regulated by the acetylation of the p65 component (Schmitz et al., 2004). SIRT1
can interact with RelA/p65 protein in the NF-B complex and specifically
deacetylates lysine 310, which has been demonstrated to potentiate the
transactivation capacity of the NFB complex (Chen and Greene, 2003). Several
studies have indicated that SIRT1 is a potent inhibitor of NF-B transcription
(Yeung et al., 2004; Chen et al., 2005b). The signaling link between SIRT1 and
NF-B is especially interesting with respect to aging, as a consequence of the
release of the SIRT1 brake, the transactivation efficiency of NF-B factor is
potentiated, which evokes immune activation and inflamm-aging.
Aging process is also regulated by autophagy. It has been identified the signaling
pathways that regulate autophagic degradation and SIRT1 is a potent regulator
of autophagic degradation (Lee et al., 2008), SIRT1 can interact with and
deacetylates several components in the complexes of forming autophagosomes,
such as Atg5, Atg7, and Atg8 proteins (Lee et al., 2008). There is a clear overlap
between the signaling networks regulating both aging and autophagocytosis,
which emphasizes the important role of autophagy in the regulating of aging and
age-related degenerative diseases. It is evident that increase in autophagy can
extend lifespan. mTOR activity is suppressed by CR, reduction in mTOR signaling
is a logic candidate mechanism for the anti-aging benefits of CR. Through
deacetylation of a variety of proteins involved in autophagy process, SIRT1 can
regulate physiological process during aging and moderated by CR (Haigis and
Guarente, 2006).

Efficient control of energy metabolic homeostasis is a hallmark of improved

healthspan and extended lifespan. The AMPK and SIRT1 signaling pathways are
highly conserved energy sensor of increased levels of AMP and NAD +,
respectively, AMPK signaling is involved in the regulation of energy metabolic
homeostasis (Hardie, 2011). Canto and Auwerx (2009) demonstrated that the
activation of AMPK stimulated the functional activity of SIRT1 by increasing the
intracellular concentration of NAD+. Interestingly, SIRT1 was able to deacetylate
LKB1 kinase which subsequently increased its activity (Lan et al., 2008). Since
LKB1 is an upstream activator of AMPK, this signaling pathway stimulates the
activation of AMPK. This positive feedback loop between SIRT1 and AMPK can
also potentiate the function of the other AMPK-activated signaling pathways. The
close relationship between AMPK and SIRT1 is evidence that energy balance
effectively controls cellular responses via an integrated signaling network. AMPK
can inhibit the activity of mTOR complex via two different mechanisms, either by
directly phosphorylating the Raptor, a regulatory component of mTORC1, or by
the phosphorylation of tuberous sclerosis protein 2 (TSC2), which subsequently
suppresses the activity of mTOR (Jung et al., 2010; Mihaylova and Shaw, 2011).
Taken together, SIRT1 interacts with other key anti-aging signaling pathways
thereby contributing to longevity control.

It has been established that aging is a known risk factor for many
neurodegenerative diseases including Alzheimers disease (AD), Parkinsons
disease (PD), Wallerian neurodegeneration, Huntingtons disease (HD), and
amyotrophic lateral sclerosis (ALS). The pathomechanisms involved in these
disorders involve common biochemical pathways and processes, including
protein misfolding, oligomerization, and aggregation, proteolysis, post-
translational modifications, mitochondrial dysfunction, abnormal metabolic
processes, and proinflammatory and proapoptotic responses that we discuss in
the next section.

SIRT1 and Age-Associated Neurological Diseases

Wallerian degeneration
Wallerian degeneration refers to axonal death and degradation after focal injury,
followed by breakdown of myelin sheath. The neuroprotective effect of SIRT1
against Wallerian degeneration was first discovered in wlds transgenic mice
(Perry et al., 1990). These mice exhibited a significant delay in axonal
degeneration after physical or chemical injury. The mechanistic basis for the
delayed axonal damage was apparently associated with the mutant wlds
chimeric protein. It has been shown that Nicotinamide mononucleotide
adenylyltransferase 1 (NMNAT-1) activity plays an important role in the
prevention of axonal damage, exerting its protective effects through SIRT1
activation, as the neuroprotection is blocked by the SIRT1 inhibitor sirtinol or
siRNA-mediated SIRT1 silencing (Araki et al., 2004; Sasaki et al., 2009; Babetto
et al., 2010). The role of SIRT1 remains controversial, however, as both SIRT1-
dependent (Araki et al., 2004) and -independent mechanisms are reported (Wang
et al., 2005b).

Alzheimers disease

Alzheimers disease is a terminal neurodegenerative disease, causing neuronal

death and brain atrophy. The pathological hallmarks of AD are the intracellular
tangles and extracellular plaques in brain. The tangles, also known as
neurofibrillary tangles, are formed by accumulation of insoluble tau proteins, and
the plaques are deposits of -amyloid (A) peptides, typically consisting of 4042
amino acid residues.

The protective effect of SIRT1 against AD was initially observed in CR studies,

where CR reduced A and plaque generation in the brains of transgenic AD mice
(Patel et al., 2005; Wang et al., 2005a). Similarly, the reduction of A was also
noticed in the cortex of fasted squirrel monkeys and is inversely correlated with
SIRT1 levels (Qin et al., 2006a). These studies imply that SIRT1 is involved in
neuroprotection against AD. Indeed, recent studies demonstrate that SIRT1
activation reduces the neuronal death and brain atrophy that are characteristic
of AD (Chen et al., 2005b; Qin et al., 2006b; Kim et al., 2007; Donmez et al.,
2010; Min et al., 2010). SIRT1 deficiency is associated with increased levels of
phosphorylated-tau in neurons (Min et al., 2010) and the amount of
neurofibrillary tangles in AD brains (Julien et al., 2009).

Moreover, recent studies show that either administration of resveratrol or

overexpression of SIRT1 reduces A levels both in vitro and in vivo (Chen et al.,
2005b; Qin et al., 2006b; Donmez et al., 2010). SIRT1 overexpression stimulates
the production of -secretase in neurons and mice by two pathways: activating
the retinoic acid receptor (RAR; Donmez et al., 2010) and inhibiting the rho-
associated, coiled-coil-containing protein kinase 1 (ROCK1; Qin et al., 2006b).
Increased levels of -secretase enhance normal process of Amyloid precursor
protein (APP), leading to decreased generation of toxic A. In addition, SIRT1 also
reduces the NF-kappaB pathway in microglia and decreases A level (Chen et al.,
2005b). Taken together, these results establish that SIRT1 protects against AD
by multiple mechanisms, including degradation of tau and reducing levels of A.

Parkinsons disease
Parkinsons disease is a common neurodegenerative disease caused by the
death of dopaminergic neurons of the substantia nigra in the midbrain. The
major symptoms of PD are rigidity, tremor, and bradykinesia. Our early study
found that CR or use of 2-deoxy-D-glucose, a glucose analog, reduces the loss of
dopaminergic neurons in mice and improves motor function, implying that SIRT1
may be involved in the protection (Duan and Mattson, 1999). The levels of SIRT1
in dopaminergic neurons are sharply decreased by treatment with neurotoxins,
such as rotenone, 6-hydroxydopamine, -synuclein, or 1-methyl-4-phenyl-
1,2,3,6-tetrahydropyridine (MPTP; Alvira et al., 2007; Pallas et al., 2008; Albani et
al., 2009), which are agents widely used to model PD. Additionally, SIRT1
overexpression (Wareski et al., 2009) or activation by resveratrol (Okawara et al.,
2007; Chao et al., 2008; Albani et al., 2009) slows neuronal death as well as
neurodegeneration in PD models both in vivo and in vitro (Donmez et al., 2012),
indicating a neuroprotective role of SIRT1 against PD. Not all studies showed a
protective role of SIRT1, however. For example, no protection was observed in an
MPTP-induced PD model in SIRT1 transgenic mice (Kakefuda et al., 2009).
Nevertheless, despite the controversy, most research demonstrates a protective
role of SIRT1 against PD, although the mechanisms are unclear.

Huntingtons disease

Huntingtons disease is an autosomal dominant hereditary disease with onset in

middle-age. It is caused by a trinucleotide repeat mutation in the huntingtin gene
that results in an increased number of glutamine residues in the N-terminus of
the huntingtin protein which causes abnormal protein aggregation and ultimately
neuronal death. Our previous study showed that CR could ameliorate the motor
phenotype and extend survival of HD mice (Duan et al., 2003), indicating that
pathways related to energy metabolism can modify disease progression in the
disease. CR increases mitochondrial biogenesis by inducing endothelial nitric
oxide synthase (eNOS), and NO can activate the SIRT1 gene (Nisoli et al.,
2005; Haigis and Guarente, 2006), which is the mammalian ortholog of yeast
Sir2, and a highly conserved NAD +-dependent protein deacetylase. Moreover,
SIRT1 has been suggested to mediate some beneficial effects of CR (Canto and
Auwerx, 2009; Wakeling et al., 2009; Shimokawa and Trindade, 2010; Chalkiadaki
and Guarente, 2012).

The first report demonstrating the connection between SIRT1 and HD came from
studies by Parker et al. (2005), who found that overexpression of Sir2.1 or
treatment with resveratrol rescued neuronal dysfunction phenotypes induced by
mutant polyglutamine in Caenorhabditis elegans. In contrast to the
neuroprotective effect of Sir2.1 in C. elegans, Pallos et al. (2008) reported that
50% reduction of Sir2 extended survival and preserved neurons containing
photoreceptor in flies expressing mutant Htt. Interestingly, in the fly model
system, overexpression of Sir2 does not reduce the lethality or the level of
neuronal degeneration caused by mutant Htt. Studies in both C.
elegans and Drosophila suggest that complete loss of Sir2 is deleterious in the
worm (Parker et al., 2005) and is deleterious compared with heterozygous loss in
mutant Htt-challenged flies (Pallos et al., 2008). Although heterozygous loss of
Sir2 is protective in flies, heterozygous loss of Sir2 in worms was not examined.
Nevertheless, reduction of Sir2 neither alters the life-span of flies not expressing
Htt nor siblings expressing Htt. Several aspects of the role of sirtuins in lifespan
in C. elegans and Drosophila are controversial, and studies have indicated that
Sir2 overexpression did not increase lifespan and that dietary restriction
increased lifespan in flies independently of dSir2 (Burnett et al., 2011).
Nonetheless, overexpression of Sir2 increases the longevity of normal flies and
the longevity of diseased flies is slightly increased by elevated Sir2 (Pallos et al.,
2008). The different results might be due to the amount of Sir2, its activation
status, and different downstream targets involved. These controversial results
warrant further investigation of the role of SIRT1 in mammalian systems.

Indeed, two independent studies by our group (Jiang et al., 2011) and Kraincs
group (Jeong et al., 2011) demonstrated that modulating the levels of SIRT1 has
therapeutic benefit in three different HD mouse models, and putative
downstream targets of SIRT1 involved in improved disease outcomes are also
identified. These two studies provide compelling support to the view that SIRT1
provides beneficial effects in HD mouse models, but also raise important
questions. It is possible that the contradictory results on the effects of SIRT1 in
models of HD might be explained by different effector pathways or mechanisms
and by context-dependent effects or different levels of SIRT1 activation. SIRT1
has numerous targets, and different models of HD display different phenotypes
by activating various targets and mechanisms. Therefore, it is not surprising to
observe contradictory data, especially in different species and different models.

Amyotrophic Lateral Sclerosis

Amyotrophic lateral sclerosis is a chronic, fatal neurodegenerative disease,

characterized pathologically by the death of motor neurons in the spinal cord and
cortex, possibly induced by a deficiency in the enzyme superoxide dismutase 1
(SOD1; Rosen, 1993). In the animal model of ALS where a mutant form of SOD1
is expressed, SIRT1 levels are upregulated in motor neurons (Kim et al., 2007).
SIRT1 overexpression protects neurons against toxicity induced by the mutant
SOD1 in both cultured neurons and mouse brain (Kim et al., 2007). This
protection corresponds to the increased deacetylation of p53. Resveratrol also
enhances the protective effect of SIRT1 in a mouse model of ALS (Kim et al.,
2007; Markert et al., 2010), but multiple doses are appear to be necessary to
improve neurological function and increase the longevity of mice (Markert et al.,
2010).

Multiple sclerosis

Multiple sclerosis is a myelin sheath disease with lesions typically located in the
brain, spinal cord or cranial nerves, and, most commonly, in the optic nerve. The
causes of multiple sclerosis are not fully identified but likely arise from an
autoimmune etiology; therefore, it is traditionally treated as an inflammatory
disease. Recently, however, multiple sclerosis has also been considered to be a
neurodegenerative disease because of the co-existence of permanent axonal
damage, neuronal loss, and neurological disability in patients with the disease
(Lassmann, 2010; Shindler et al., 2010). In a mouse model of multiple sclerosis,
experimental autoimmune encephalomyelitis (EAE), SIRT1 activation by SRT501
or SRT1720 maintains axonal density, prevents neuronal loss, and improves
neuronal dysfunction (Shindler et al., 2007, 2010). SIRT1 inhibition with Sirtinol
attenuates the neuroprotective effects of SRT501 (Shindler et al., 2010),
suggesting a protective role of SIRT1 in multiple sclerosis. However, further
investigations are necessary to fully delineate the role of SIRT1 in multiple
sclerosis.

Cerebral ischemia

Ischemic stroke is a common neurological disease caused by the sudden

reduction or cessation of blood flow to the brain, leading to infarction. The
clinical management of stroke is difficult and current drugs must be administered
within a limited time window after the onset of the stroke to provide clinical
benefit. Promising candidates for neuroprotective strategies include
preconditioning, mild hypothermia, and the use of chemical and biological
compounds targeting critical molecular mediators of neuronal death and survival.

The neuroprotective effect of SIRT1 was first reported in ischemic preconditioning

and the SIRT1 activating compound resveratrol reduced neuronal injury of the
hippocampus in global cerebral ischemia in rats. Increased SIRT1 activity was
also shown to be a common mechanism for the protective effects of
preconditioning and resveratrol (Raval et al., 2006; Morris et al., 2011). Sirtinol,
an inhibitor of SIRT1 activity, abolished the neuroprotection of preconditioning
and resveratrol (Raval et al., 2006), indicating that SIRT1 plays a key role in
mediating neuroprotection. This neuroprotective role is further supported by two
recent studies (Chong and Maiese, 2008; Della-Morte et al., 2009) showing that
SIRT1 activation reduces ischemic neuronal injuries.

Another study showed that, in primary neuronal culture, pretreatment with

NAD+ pretreatment markedly reduces neuronal death induced by oxygen-glucose
deprivation, an in vitro model of ischemia (Wang et al., 2008). SIRT1 is necessary
for NAD+ neuroprotection, as NAD+ treatment upregulates SIRT1 expression and
activity, and SIRT1 knockdown attenuates the protection mediated by
NAD+ (Wang et al., 2008). NAMPT overexpression reduces ischemic infarct,
whereas NAMPT inhibition aggravates ischemic injuries. The protective effect of
NAMPT is SIRT1-dependent, as SIRT1 knockout blocks the protection (Wang et al.,
2011).

Despite the aforementioned evidence, controversy exists over the protective

effect of SIRT1 against ischemia. In a study with SIRT1 transgenic mice, where
human SIRT1 was overexpressed under the control of rat neuron-specific enolase
promoter, no neuroprotection was observed against stroke as SIRT1 and wild-
type mice demonstrated almost indistinguishable infarct volumes and
neurological deficiency scores (Kakefuda et al., 2009). The discrepancy between
this study and the others was probably due to the sustained high level of SIRT1,
because it may consume too much or even deplete NAD +, which could
exaggerate neuronal injury (Wang et al., 2008; Kakefuda et al., 2009; Liu et al.,
2009). Therefore, it is possible that NAD+ deficiency compromised the
neuroprotective effect of SIRT1. In another study, nicotinamide, a compound that
inhibits SIRT1 action, showed neuroprotection against ischemic injury, implying
that SIRT1 might have a detrimental effect against stroke (Chong et al., 2005).
However, this report might overlook other functions of nicotinamide, including
that of precursor for NAD+ synthesis. In fact, the same group later reported that
SIRT1 overexpression prevents neurons from apoptosis after oxidative stress
(Chong and Maiese, 2008).

SIRT1 in Clinical Practice

SIRT1-activating compounds have not yet been proven to be clinically useful for
the treatment of neurodegenerative diseases. Preclinical studies have been
performed in various neurodegenerative disease models, however. The
information obtained from such studies could prove to be valuable for designing
SIRT1-activating molecules that may be more likely to be useful clinically. The
discovery of such molecules is becoming increasingly important, considering the
limitations of genetic manipulations and the lack of unequivocal evidence of
specific SIRT1 activation by prototype molecules like resveratrol (Sauve, 2009).

Several rational strategies based on the available protein structure and the
catalytic pathways have been designed to develop small molecules that
selectively activate sirtuins (Sauve, 2009). One strategy involves designing
resveratrol-like molecules, which has not yielded successful results as the in
vivo mechanism of SIRT1 activation is not fully understood. Another approach
aims to increase the cellular levels of NAD+ in order to activate SIRT1 function.
This approach has the advantage of harnessing a natural metabolic pathway to
enhance SIRT1 functions. Moreover, naturally occurring metabolites present the
least risk of toxicity. The efficacies of agents that have been used to enhance
NAD+ are still questionable, however, and NAD + enhancement affects a host of
other physiological pathways, so that the approach is not specific to SIRT1. A
third strategy currently in the proof-of-principle stage designated nicotinamide
derepression is based on countering the inhibitory effect of nicotinamide on
sirtuins by designing molecules that are antagonistic to nicotinamide. This
approach is still in its infancy and has not provided compounds with desired
potency, but is an attractive strategy to develop further. Details of efforts to
discover SIRT1-activating molecules have recently been comprehensively
reviewed by Blum et al. (2011).

Future Perspectives on SIRT1

Over the last decade, our understanding of the biology of sirtuins has vastly
increased from its original description as an NAD +-dependent class III histone
deacetylase that can control the lifespan of yeast. Of particular interest is the
discovery that SIRT1 not only deacetylates histones, but also some well-known
transcriptional regulators, thereby modulating a wide array of biological
processes. An exciting aspect is that SIRT1 mediates neuroprotection against
both acute and chronic neurological diseases. Importantly, SIRT1 activity is
enhanced by small-molecule compounds; therefore, development of small-
molecule activators could lead to novel therapies against neurological diseases.
One of the broad protective mechanisms of SIRT1 is to suppress genome-wide
gene transcription via histone deacetylation. Furthermore, SIRT1 selectively
suppresses genes involved in fat storage, apoptosis, and inflammation. Adding to
the complexity of SIRT1-mediated cell survival, SIRT1 specifically promotes the
transcription of a set of genes related to cell survival, energy metabolism, and
mitochondrial biogenesis. SIRT1 thus has multifaceted mechanisms with the end
goal to increase cell viability.

Although extensively studied, the biological functions of SIRT1 remain only

partially characterized. With respect to mechanism of action, there are several
substantial unknowns. For example, it is not known how SIRT1 specifically
increases transcription of beneficial genes while it simultaneously suppresses
universal transcription. It would be of interest to determine whether similar
mechanisms exist for genes upregulated by SIRT1-mediated activation of
transcription. Another issue is the paradoxical effect of SIRT1. For example,
whereas SIRT1 directly suppresses PPAR transcriptional activity directly (Picard
et al., 2004), it also activates PGC-1 (Nemoto et al., 2005; Rodgers et al., 2005),
which could increase the transcriptional activities of PPAR (Puigserver et al.,
1998).

Although SIRT1 has been found to be neuroprotective in numerous studies, it is

clear from the diverse pathological mechanisms manifested in
neurodegenerative disorders that the role of SIRT1 requires more detailed study.
The availability of crystal structures and detailed mechanistic analysis are helpful
in discovering SIRT1 modulators, but would be of limited value if they failed to
reach clinical trials, thus emphasizing the importance of developing robust
animal models for investigating molecular mechanisms involved in SIRT1
activation. Moreover, the potential negative effects of SIRT1 activation and
energy depletion need further investigation in animal models. The clinical
success of sirtuin activating compounds (STACs) in neurodegenerative diseases
relies overwhelmingly on developing new strategies and designing molecules
based on the sirtuin chemistry and molecular pathways activated by SIRT1. A
very recent study performed by Hubbard et al. (2013) demonstrated that the
specific hydrophobic motifs in SIRT1 substrates such as PGC-1 and FOXO3a
facilitate SIRT1 activation by STACs, implying that SIRT1 can be directly activated
through an allosteric mechanism common to chemically diverse STACs. In
summary, there is no doubt that SIRT1 holds promising therapeutic potential for
neurodegenerative disorders.

SIRT3, Energy Metabolism and Aging

This section summarizes the studies on the role of mitochondrial SIRT3 in energy
metabolism and protection against oxidative stress and age-associated
dysfunction (Figure 2).

FIGURE 2
FIGURE 2. Schematic overview of molecular targets of SIRT3 and the
role in regulating metabolism in mitochondria.

SIRT3 and Mitochondria

Mitochondria are not only the powerhouse for ATP production but also the main
sites where reactive oxygen species (ROS) are generated and the intrinsic
apoptotic signaling pathway is initiated (Salminen et al., 2008b). The functions of
mitochondrial proteins are altered when they are deacetylated by NAD +-
dependent mitochondrial deacetylases, including SIRT3, SIRT4, and SIRT5. All
mitochondrial sirtuins are present in the mitochondrial matrix (Howitz et al.,
2003; Rogina and Helfand, 2004). Since mitochondria contain their own DNA,
transcription factors, histone-like proteins, and protein synthesis systems,
mitochondrial sirtuins deacetylate a set of targets within the mitochondria that
are distinct from those of nuclear proteins (Tissenbaum and Guarente,
2001; Wood et al., 2004). Although precise mechanistic information is still
lacking, evidence is emerging to suggest that mitochondrial sirtuins protect
against oxidative stress (Braunstein et al., 1993; Baur et al., 2006).

Among the mitochondrial sirtuins, SIRT3 functions have been characterized in the
greatest detail. Initial studies of SIRT3-deficient mice indicated that loss of SIRT3,
but not SIRT4 or SIRT5, led to dramatic protein hyperacetylation within
mitochondria, suggesting that SIRT3 is the major mitochondrial deacetylase
activity (Dali-Youcef et al., 2007). In humans, full-length SIRT3 is a 44-kD protein
with an N-terminal mitochondrial targeting sequence that is an enzymatically
inactive in vitro. It is proteolytically processed in mitochondria to a mature 28 kD
catalytically active deacetylase (Onyango et al., 2002; Schwer et al., 2002). The
first mouse SIRT3 cDNA sequence identified encoded a 28-kD protein lacking the
N-terminal mitochondrial targeting sequence (Yang et al., 2000). Several recent
studies have identified a longer isoform of murine SIRT3 encoding a 37 kD
protein, however, that can be imported into mitochondria and processed into the
mature 28 kD protein (Cooper et al., 2009; Jin et al., 2009; Bao et al., 2010; Yang
et al., 2010). Whether or not an active fraction of SIRT3 exists outside
mitochondria and modifies extra-mitochondrial proteins remains controversial.

SIRT3 and Metabolic Homeostasis

Emerging data have shown that one major function of SIRT3 is regulation of
mitochondrial electron transport chain activity to maintain energy homeostasis.
The main energy source in mitochondria is pyruvate, a product of glycolysis.
Alternatively, mitochondria also burn fatty acids, amino acids, and acetates when
pyruvate is deficient. For fatty acid catabolism, long-chain acyl coenzyme A
dehydrogenase (LCAD) is a key enzyme that breaks down fatty acids and
generates acetyl-CoA, stimulating -oxidation. In SIRT3 knockout mice, LCAD is
hyperacetylated at Lys42, leading to decreases in enzymatic activity, -
oxidation, and ATP level (Hirschey et al., 2010). Interestingly, these mice do not
tolerate cold exposure during fasting (Hirschey et al., 2010). SIRT3 directly
deacetylates LACD at Lys42 and increases LACD activity (Hirschey et al., 2010).
In addition, SIRT3 may promote -oxidation via multiple mechanisms, such as by
deacetylating other -oxidation enzymes, including the short-chain L-3-
hydroxyacyl-CoA dehydrogenase and the very-long-chain acyl coenzyme A
dehydrogenase (Hallows et al., 2011), facilitating mitochondrial adaptation to
fuel changes.

Glucose is the major energy source for cells. When its availability is limited,
however, alternative fuels become increasingly important for cell survival. The
first step of glycolysis is the conversion of glucose to glucose-6-phosphate, a
reaction catalyzed by hexokinases. It is reported that SIRT3 deacetylates
cyclophilin D (Hafner et al., 2010; Shulga et al., 2010), which leads to the
dissociation of hexokinase II and mitochondria, decreases glucose metabolism,
and stimulates oxidative phosphorylation (Shulga et al., 2010).

Acetate derived from acetic acid and alcohol is also used as a mitochondrial fuel,
although this only occurs in extreme circumstances of nutrient depletion. In the
initial step acetate is converted to acetyl-CoA catalyzed by acetyl-CoA
synthetases. Acetyl-CoA synthetase 2 is the mitochondrial form of the enzyme.
SIRT3 deacetylates acetyl-CoA synthetase 2 and enhances its activity, leading to
increased production of acetyl-CoA (Hallows et al., 2006; Schwer et al., 2006).
The acetyl-CoA from fatty acids and acetates as well as -ketoglutarate from
amino acids can enter the Krebs cycle. These two reactions are enhanced by
SIRT3 (Hallows et al., 2006; Schwer et al., 2006; Lombard et al., 2007; Schlicker
et al., 2008). Additionally, SIRT3 directly stimulates the Krebs cycle. The third
step of the cycle is the conversion of 6-carbon isocitrate to 5-carbon -
ketoglutarate, a process catalyzed by isocitrate dehydrogenase 2 (IDH2). A
recent study shows that SIRT3 directly deacetylates this dehydrogenase to
increase its activity (Someya et al., 2010).

NADH dehydrogenase 1 alpha subcomplex subunit 9 (NDUFA9) is an enzyme of

mitochondrial complex I that is acetylated at Lys370 (Kim et al., 2006). SIRT3
physically interacts with NDUFA9 and deacetylates it. SIRT3 knockout enhances
its acetylation and reduces the activity of complex I (Ahn et al., 2008), indicating
that SIRT3 is a positive regulator of complex I. Complex II, also known as
succinate dehydrogenase, is composed of four subunits, including the
flavoprotein succinate dehydrogenase subunit A (SdhA). In SIRT3 knockout mice,
SdhA is hyperacetylated at several lysine residues, and shows decreased activity
of complex II (Cimen et al., 2010). SIRT3 overexpression reverses the acetylation
of SdhA and increases complex II activity (Cimen et al., 2010), indicating that
SdhA is a SIRT3 substrate, and that SIRT3 is also a positive regulator of complex
II.
SIRT3 is also reported to bind the alpha subunit 1 of the F1 particle of ATP
synthase (Law et al., 2009), but the function is unclear. Taken together, these
results suggest that SIRT3 promotes ATP generation through enhancing action of
several enzymes involved in energy metabolism. Further supporting this role,
SIRT3-knockout mice show substantial acetylation of mitochondrial proteins, and
have reduced ATP levels at baseline and during cellular stress (Ahn et al., 2008).

Mitochondria are also the major sites for the generation of the ROS, superoxide,
and also where the superoxide is dismuted by mitochondrial MnSOD. Recent
reports show that SIRT3 deacetylates MnSOD at Lys122 and increases its activity,
reducing oxidative and radiation stress in mice (Qiu et al., 2010; Tao et al., 2010).
Overexpression of SIRT3 protects HEK293 from oxidative stress and prevents
age-related cochlear cell death in mice (Someya et al., 2010). Overall, this
suggests anti-oxidative and neuroprotective roles of SIRT3. New data suggest
that the SIRT3 deacetylase plays a key role in bolstering mitochondrial anti-
oxidant defenses during CR (Qiu et al., 2010; Someya et al., 2010).

In future studies, it will be of interest to define the mechanism whereby

acetylation of electron transport chain subunits affects generation of ATP. It is
also important to elucidate why it might be desirable under some physiologic
conditions to downregulate electron transport chain activity by increased
acetylation. As an added wrinkle, SIRT3 negatively regulates translation within
mitochondria by deacetylating the ribosomal protein MRPL10, a function
proposed to reduce respiration (Yang et al., 2010).

SIRT3 levels are increased in adipose tissue, skeletal muscle, and liver during CR
(Shi et al., 2005; Palacios et al., 2009; Schwer et al., 2009; Hirschey et al., 2010),
and conversely decline in response to high-fat feeding (Palacios et al., 2009; Bao
et al., 2010; Kendrick et al., 2011). These expression data suggest that SIRT3
might play a role in the response to metabolic homeostasis. In mammals, two
independent studies showed that SIRT3 interacts with and deacetylates acetyl-
CoA synthetase 2 (AceCS2) at the active site lysine to promote AceCS2 activity
(Hallows et al., 2006; Schwer et al., 2006). Under fed conditions, the majority of
acetyl-CoA is generated through metabolism of pyruvate by Pyruvate
dehydrogenase complex (PDC) and by fatty acid -oxidation, largely bypassing
the need for AceCS2. In this regard, studies of AceCS2-deficient mice revealed
that AceCS2 is specifically required for metabolic homeostasis when the mice are
fed a low carbohydrate/high fat diet (LC/HFD); AceCS2-deficient animals are
essentially normal on a chow diet but show poor weight gain, hypothermia,
hypoglycemia, and impaired survival on an LC/HFD (Sakakibara et al., 2009).
Presumably the role of SIRT3 in regulating AceCS2 could also be important during
fasting, when acetate can be used as a source of energy in extrahepatic tissues
(Hirschey et al., 2010). Indeed, SIRT3 has recently been shown to deacetylate
and activate 3-hydroxy-3-methylglutaryl-CoA synthase 2 (HMGCS2), a
mitochondrial enzyme that converts acetyl-CoA into ketone bodies
(acetoacetate, -hydroxybutyrate, and acetone) in the liver under fasting
conditions, which can in turn be used as a source of energy in certain tissues
including the brain (Shimazu et al., 2010). SIRT3-deficient mice are unable to
produce normal levels of ketone bodies upon fasting.

SIRT, Lifespan and Age-Associated Phenotypes

The role of SIRT3 in aging is of considerable interest because it appears to

suppress ROS, one of the causes contributing to the process of aging. In addition
to elucidating its roles in regulating specific biochemical pathways in
mitochondria, there is great interest in testing whether SIRT3 might modulate
age-associated phenotypes, or indeed lifespan itself. In this regard, some studies
have linked polymorphisms in the SIRT3 genomic locus to human longevity,
although others have failed to demonstrate this association (Rose et al.,
2003; Bellizzi et al., 2005, 2007; Lescai et al., 2009). A polymorphism associated
with decreased SIRT3 mRNA expression was present in cohorts of young but not
old men, suggesting that reduced SIRT3 expression may be detrimental to
survival in old age (Bellizzi et al., 2005, 2009). In sedentary individuals, SIRT3
protein expression declined with age in skeletal muscle mitochondria,
concomitant with a reduction in respiratory function (Lanza et al., 2008).

Age-related hearing loss (ARHL) is a common problem in the elderly, occurring

secondary to cell loss and other degenerative changes in the cochlea. An elegant
study has firmly established a role for SIRT3 in preventing ARHL (Someya et al.,
2010). One mechanism by which SIRT3 mediates this effect is by deacetylation of
IDH2 (Schlicker et al., 2008; Someya et al., 2010), which converts isocitrate to -
ketoglutarate concomitant with reduction of NADP +. NADPH in turn allows
regeneration of reduced glutathione to promote mitochondrial oxidative defense.
In response to CR, wild-type mice, but not SIRT3-deficient animals, show
increased NADPH levels, increased reduced glutathione in mitochondria, and
decreased DNA damage in the cochlea and in other tissues. In tissue culture
cells, overexpression of SIRT3 or IDH2 protects against oxidative stress-induced
cell death, and the two proteins together have a synergistic pro-survival effect.
These results do not rule out the possibility that SIRT3 might modify other
substrates in addition to IDH2 to prevent AHRL during CR. Similarly, Qiu et al.
(2010) reported that SIRT3-deficient mice fail to suppress ROS levels and
macromolecular damage during CR. They find that SIRT3 directly deacetylates
SOD2 to increase its activity during CR, whereas SIRT3-deficient mice do not
show SOD2 deacetylation in response to this diet (Qiu et al., 2010). Overall,
these papers point to crucial role for SIRT3 in suppressing oxidative damage and
its negative sequel during CR. It remains to be seen how SIRT3, or the other
mitochondrial sirtuins, might affect other phenotypes of aging and/or effects of
CR. The reduction of serum insulin and triglycerides that normally occurs during
CR is not observed in SIRT3-deficient mice (Someya et al., 2010), implying that
SIRT3 plays additional, uncharacterized roles in the adaptation to this dietary
regimen. A recent study indicates that upregulation of SIRT3 indeed reverses
aging-associated degeneration in hematopoietic stem cells (Brown et al., 2013),
and SIRT3 may promote organismal longevity by maintaining the integrity of
tissue-specific stem cells.
SIRT3 and Mitochondrial Protein Acetylation: Unresolved Questions

Acetylation of mitochondrial proteins plays a major role in regulating functions of

this organelle. Despite the rapid progress in this area, there are still many
outstanding questions remain that will no doubt provide fruitful avenues for
research for years to come. In particular, how mitochondrial proteins are
acetylated in the first place is currently unknown. The identity of putative
mitochondrial acetyltransferases remains elusive; identification of such proteins
would represent a major step forward in this field. Alternatively, or in addition to
enzymatic acetylation within mitochondria, mitochondrial proteins could in
principle be acetylated outside this organelle, before or concomitant with
mitochondrial import or even be acetylated non-enzymatically. These latter
models would not permit rapid cycles of acetylation/deacetylation of
mitochondrial proteins to regulate target protein function in response to varied
environmental challenges. Instead, after deacetylation, restoration of acetylation
status would require new protein synthesis. Such models could be distinguished
through pulse-chase experiments assessing acetylation of newly synthesized
mitochondrial proteins before and after mitochondrial import.

Similarly, how SIRT3 activity is regulated in the mitochondria is incompletely

understood. SIRT3 requires NAD+, and therefore mitochondrial NAD+ levels play a
critically important role in governing mitochondrial SIRT3 function. Increased
NADH generation from NAD+ that occurs with HFD and leads to reduced SIRT3
function may explain the increased global mitochondrial protein acetylation
observed during this diet, as could increase levels of acetyl-CoA, the substrate
for acetyltransferases (Kendrick et al., 2011). This overall increased acetylation
may represent the net effect of increased acetyltransferase activity
superimposed upon elevated SIRT3 function; alternatively, some protein species
hyperacetylated during CR or other conditions may not be substrates for
mitochondrial SIRT3. The activity of SIRT3 and other mitochondrial sirtuins might
be influenced by other conditions in addition to NAD + levels, such as post-
translational modification or interactions with regulatory proteins.

It is currently unclear whether interventions that have an impact on acetylation

of many mitochondrial proteins SIRT3 deficiency, such as CR, HFD lead to
modification of common sets of proteins on the same lysine sites, or whether this
response is tailored to different environmental perturbations. Similarly, it remains
unclear whether the mitochondrial sirtuins share common targets and/or
functions in common. This question could be addressed in mice or cells with
mitochondrial sirtuin deficiencies or knockdowns. Given that SIRT3 deacetylates
many proteins in mitochondria as well as suppresses some age-associated
phenotypes, it will be of interest to test whether acetylation of mitochondrial
proteins changes with age, either individually or globally, and whether
prevention of this effect might have a beneficial effect on health span or even
lifespan.

In addition, whereas the functional impact of acetylation on a few individual

protein targets is clear, a global understanding of how SIRT3 affects the activity
of metabolic pathways in mitochondria, cells, tissues, and the organism overall is
still lacking. Answers to these and related questions involving SIRT3 and
mitochondrial protein acetylation will no doubt reveal novel aspects of
mitochondrial biology, and perhaps ultimately provide the basis for novel
therapeutic strategies for a variety of disorders.

Further Perspectives

In the past decade, the function of mammalian sirtuins has been investigated in
greater detail than ever before, and we now have a much better molecular
understanding of the multiple roles this unique family of enzymes plays in
seemingly every biological process. There is little doubt that sirtuins have
emerged as critical modulators of metabolic adaptive responses, and their
activities have been linked to metabolic abnormalities as well as age-associated
neurodegeneration. Yet, key questions will keep investigators busy in the coming
years. We still have poor understanding of the molecular mechanisms regulating
expression and activity of the sirtuins, and of the precise stimuli that regulate
these proteins, and whether the activities of different sirtuins are regulated in a
coordinated fashion. In other words, is there cross-talk between sirtuins? Will
future studies cement the argument that sirtuins are, indeed, critical modulators
of lifespan? This review has focused on SIRT1 and SIRT3. Even less is known
about other sirtuins. Further investigation into the targets and functions of this
unique family of sirtuins will help develop new strategies for protection against
and recovery from common aging-related neurological diseases and promote
successful aging.

Conflict of Interest Statement

The author declares that the research was conducted in the absence of any
commercial or financial relationships that could be construed as a potential
conflict of interest.

References

Adler, A. S., Sinha, S., Kawahara, T. L., Zhang, J. Y., Segal, E., and Chang, H. Y.
(2007). Motif module map reveals enforcement of aging by continual NF-kappaB
activity. Genes Dev. 21, 32443257. doi: 10.1101/gad.1588507

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Ahn, B. H., Kim, H. S., Song, S., Lee, I. H., Liu, J., Vassilopoulos, A., et al. (2008). A
role for the mitochondrial deacetylase Sirt3 in regulating energy
homeostasis. Proc. Natl. Acad. Sci. U.S.A. 105, 1444714452. doi:
10.1073/pnas.0803790105

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Albani, D., Polito, L., Batelli, S., De Mauro, S., Fracasso, C., Martelli, G., et al.
(2009). The SIRT1 activator resveratrol protects SK-N-BE cells from oxidative
stress and against toxicity caused by alpha-synuclein or amyloid-beta (142)
peptide. J. Neurochem. 110, 14451456. doi: 10.1111/j.1471-4159.2009.06228.x

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Alvira, D., Yeste-Velasco, M., Folch, J., Verdaguer, E., Canudas, A. M., Pallas, M., et
al. (2007). Comparative analysis of the effects of resveratrol in two apoptotic
models: inhibition of complex I and potassium deprivation in cerebellar
neurons. Neuroscience 147, 746756. doi: 10.1016/j.neuroscience.2007.04.029

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Araki, T., Sasaki, Y., and Milbrandt, J. (2004). Increased nuclear NAD biosynthesis
and SIRT1 activation prevent axonal degeneration. Science 305, 10101013. doi:
10.1126/science.1098014

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Babetto, E., Beirowski, B., Janeckova, L., Brown, R., Gilley, J., Thomson, D., et al.
(2010). Targeting NMNAT1 to axons and synapses transforms its neuroprotective
potency in vivo. J. Neurosci. 30, 1329113304. doi: 10.1523/JNEUROSCI.1189-
10.2010

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Bao, J., Scott, I., Lu, Z., Pang, L., Dimond, C. C., Gius, D., et al. (2010). SIRT3 is
regulated by nutrient excess and modulates hepatic susceptibility to
lipotoxicity. Free Radic. Biol. Med. 49, 12301237. doi:
10.1016/j.freeradbiomed.2010.07.009

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Baur, J. A., Pearson, K. J., Price, N. L., Jamieson, H. A., Lerin, C., Kalra, A., et al.
(2006). Resveratrol improves health and survival of mice on a high-calorie
diet. Nature 444, 337342. doi: 10.1038/nature05354

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Bellizzi, D., Covello, G., Di Cianni, F., Tong, Q., and De Benedictis, G. (2009).
Identification of GATA2 and AP-1 activator elements within the enhancer VNTR
occurring in intron 5 of the human SIRT3 gene. Mol. Cells 28, 8792. doi:
10.1007/s10059-009-0110-3

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Bellizzi, D., Dato, S., Cavalcante, P., Covello, G., Di Cianni, F., Passarino, G., et al.
(2007). Characterization of a bidirectional promoter shared between two human
genes related to aging: SIRT3 and PSMD13. Genomics 89, 143150. doi:
10.1016/j.ygeno.2006.09.004

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Bellizzi, D., Rose, G., Cavalcante, P., Covello, G., Dato, S., De Rango, F., et al.
(2005). A novel VNTR enhancer within the SIRT3 gene, a human homologue of
SIR2, is associated with survival at oldest ages. Genomics 85, 258263. doi:
10.1016/j.ygeno.2004.11.003

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Biessels, G. J., and Kappelle, L. J. (2005). Increased risk of Alzheimers disease in

Type II diabetes: insulin resistance of the brain or insulin-induced amyloid
pathology? Biochem. Soc. Trans. 33, 10411044. doi: 10.1042/BST20051041

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Blum, C. A., Ellis, J. L., Loh, C., Ng, P. Y., Perni, R. B., and Stein, R. L. (2011). SIRT1
modulation as a novel approach to the treatment of diseases of aging. J. Med.
Chem. 54, 417432. doi: 10.1021/jm100861p

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Boveris, A., and Navarro, A. (2008). Brain mitochondrial dysfunction in

aging. IUBMB Life 60, 308314. doi: 10.1002/iub.46

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Braunstein, M., Rose, A. B., Holmes, S. G., Allis, C. D., and Broach, J. R. (1993).
Transcriptional silencing in yeast is associated with reduced nucleosome
acetylation. Genes Dev. 7, 592604. doi: 10.1101/gad.7.4.592

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Brooks, C. L., and Gu, W. (2009). How does SIRT1 affect metabolism, senescence
and cancer? Nat. Rev. Cancer 9, 123128. doi: 10.1038/nrc2562

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Brown, K., Xie, S., Qiu, X., Mohrin, M., Shin, J., Liu, Y., et al. (2013). SIRT3 reverses
aging-associated degeneration. Cell Rep. 3, 319327. doi:
10.1016/j.celrep.2013.01.005

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Burnett, C., Valentini, S., Cabreiro, F., Goss, M., Somogyvari, M., Piper, M. D., et
al. (2011). Absence of effects of Sir2 overexpression on lifespan in, C.
elegans and Drosophila. Nature 477, 482485. doi: 10.1038/nature10296

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Campisi, J. (2005). Suppressing cancer: the importance of being

senescent. Science 309, 886887. doi: 10.1126/science.1116801

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Canto, C., and Auwerx, J. (2009). Caloric restriction, SIRT1 and longevity. Trends
Endocrinol. Metab. 20, 325331. doi: 10.1016/j.tem.2009.03.008

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Chalkiadaki, A., and Guarente, L. (2012). Sirtuins mediate mammalian metabolic

responses to nutrient availability. Nat. Rev. Endocrinol. 8, 287296. doi:
10.1038/nrendo.2011.225

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Chao, J., Yu, M. S., Ho, Y. S., Wang, M., and Chang, R. C. (2008). Dietary
oxyresveratrol prevents parkinsonian mimetic 6-hydroxydopamine
neurotoxicity. Free Radic. Biol. Med. 45, 10191026. doi:
10.1016/j.freeradbiomed.2008.07.002

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Chen, D., Steele, A. D., Lindquist, S., and Guarente, L. (2005a). Increase in
activity during calorie restriction requires Sirt1. Science 310, 1641. doi:
10.1126/science.1118357

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Chen, J., Zhou, Y., Mueller-Steiner, S., Chen, L. F., Kwon, H., Yi, S., et al. (2005b).
SIRT1 protects against microglia-dependent amyloid-beta toxicity through
inhibiting NF-kappaB signaling. J. Biol. Chem. 280, 4036440374. doi:
10.1074/jbc.M509329200

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Chen, L. F., and Greene, W. C. (2003). Regulation of distinct biological activities of

the NF-kappaB transcription factor complex by acetylation. J. Mol. Med. (Berl.) 81,
54957. doi: 10.1007/s00109-003-0469-0

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Chong, Z. Z., Lin, S. H., Li, F., and Maiese, K. (2005). The sirtuin inhibitor
nicotinamide enhances neuronal cell survival during acute anoxic injury through
AKT, BAD, PARP, and mitochondrial associated anti-apoptotic pathways. Curr.
Neurovasc. Res. 2, 271285. doi: 10.2174/156720205774322584

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Chong, Z. Z., and Maiese, K. (2008). Enhanced tolerance against early and late
apoptotic oxidative stress in mammalian neurons through nicotinamidase and
sirtuin mediated pathways. Curr. Neurovasc. Res. 5, 159170. doi:
10.2174/156720208785425666

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Cimen, H., Han, M. J., Yang, Y., Tong, Q., Koc, H., and Koc, E. C. (2010). Regulation
of succinate dehydrogenase activity by SIRT3 in mammalian
mitochondria. Biochemistry 49, 304311. doi: 10.1021/bi901627u

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Cooper, H. M., Huang, J. Y., Verdin, E., and Spelbrink, J. N. (2009). A new splice
variant of the mouse SIRT3 gene encodes the mitochondrial precursor
protein. PLoS ONE 4:e4986. doi: 10.1371/journal.pone.0004986

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Dali-Youcef, N., Lagouge, M., Froelich, S., Koehl, C., Schoonjans, K., and Auwerx, J.
(2007). Sirtuins: the magnificent seven, function, metabolism and
longevity. Ann. Med. 39, 335345. doi: 10.1080/07853890701408194

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Della-Morte, D., Dave, K. R., DeFazio, R. A., Bao, Y. C., Raval, A. P., and Perez-
Pinzon, M. A. (2009). Resveratrol pretreatment protects rat brain from cerebral
ischemic damage via a sirtuin 1-uncoupling protein 2
pathway. Neuroscience 159, 9931002. doi: 10.1016/j.neuroscience.2009.01.017

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Dietrich, M. O., Antunes, C., Geliang, G., Liu, Z. W., Borok, E., Nie, Y., et al. (2010).
Agrp neurons mediate Sirt1s action on the melanocortin system and energy
balance: roles for Sirt1 in neuronal firing and synaptic plasticity. J. Neurosci. 30,
1181511825. doi: 10.1523/JNEUROSCI.2234-10.2010

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Donmez, G., Arun, A., Chung, C. Y., McLean, P. J., Lindquist, S., and Guarente, L.
(2012). SIRT1 protects against alpha-synuclein aggregation by activating
molecular chaperones. J. Neurosci. 32, 124132. doi: 10.1523/JNEUROSCI.3442-
11.2012

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Donmez, G., Wang, D., Cohen, D. E., and Guarente, L. (2010). SIRT1 suppresses
beta-amyloid production by activating the alpha-secretase gene
ADAM10. Cell 142, 320332. doi: 10.1016/j.cell.2010.06.020

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Drew, B., and Leeuwenburgh, C. (2004). Ageing and subcellular distribution of

mitochondria: role of mitochondrial DNA deletions and energy production. Acta
Physiol. Scand. 182, 333341. doi: 10.1111/j.1365-201X.2004.01371.x

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Duan, W., Guo, Z., Jiang, H., Ware, M., Li, X. J., and Mattson, M. P. (2003). Dietary
restriction normalizes glucose metabolism and BDNF levels, slows disease
progression, and increases survival in huntingtin mutant mice. Proc. Natl. Acad.
Sci. U.S.A. 100, 29112916. doi: 10.1073/pnas.0536856100

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Duan, W., and Mattson, M. P. (1999). Dietary restriction and 2-deoxyglucose

administration improve behavioral outcome and reduce degeneration of
dopaminergic neurons in models of Parkinsons disease. J. Neurosci. Res. 57,
195206. doi: 10.1002/(SICI)1097-4547(19990715)57:2

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Finley, L. W., and Haigis, M. C. (2009). The coordination of nuclear and

mitochondrial communication during aging and calorie restriction. Ageing Res.
Rev. 8, 173188. doi: 10.1016/j.arr.2009.03.003

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Guarente, L., and Franklin, H. (2011). Epstein lecture: sirtuins, aging, and
medicine. N. Engl. J. Med. 364, 22352244. doi: 10.1056/NEJMra1100831

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Hafner, A. V., Dai, J., Gomes, A. P., Xiao, C. Y., Palmeira, C. M., Rosenzweig, A., et
al. (2010). Regulation of the mPTP by SIRT3-mediated deacetylation of CypD at
lysine 166 suppresses age-related cardiac hypertrophy. Aging (Albany NY) 2,
914923.

Pubmed Abstract | Pubmed Full Text

Haigis, M. C., and Guarente, L. P. (2006). Mammalian sirtuins emerging roles in

physiology, aging, and calorie restriction. Genes Dev. 20, 29132921. doi:
10.1101/gad.1467506

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Hallows, W. C., Lee, S., and Denu, J. M. (2006). Sirtuins deacetylate and activate
mammalian acetyl-CoA synthetases. Proc. Natl. Acad. Sci. U.S.A. 103, 10230
10235. doi: 10.1073/pnas.0604392103

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Hallows, W. C., Yu, W., Smith, B. C., Devries, M. K., Ellinger, J. J., Someya, S., et al.
(2011). Sirt3 promotes the urea cycle and fatty acid oxidation during dietary
restriction. Mol. Cell. 41, 139149. doi: 10.1016/j.molcel.2011.01.002

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Hardie, D. G. (2011). AMP-activated protein kinase: an energy sensor that
regulates all aspects of cell function. Genes Dev. 25, 18951908. doi:
10.1101/gad.17420111

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Herranz, D., Munoz-Martin, M., Canamero, M., Mulero, F., Martinez-Pastor, B.,
Fernandez-Capetillo, O., et al. (2010). Sirt1 improves healthy ageing and protects
from metabolic syndrome-associated cancer. Nat. Commun. 1, 3. doi:
10.1038/ncomms1001

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Hirschey, M. D., Shimazu, T., Goetzman, E., Jing, E., Schwer, B., Lombard, D. B.,
et al. (2010). SIRT3 regulates mitochondrial fatty-acid oxidation by reversible
enzyme deacetylation. Nature 464, 121125. doi: 10.1038/nature08778

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Howitz, K. T., Bitterman, K. J., Cohen, H. Y., Lamming, D. W., Lavu, S., Wood, J. G.,
et al. (2003). Small molecule activators of sirtuins extend Saccharomyces
cerevisiae lifespan. Nature 425, 191196. doi: 10.1038/nature01960

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Hubbard, B. P., Gomes, A. P., Dai, H., Li, J., Case, A. W., Considine, T., et al. (2013).
Evidence for a common mechanism of SIRT1 regulation by allosteric
activators. Science 339, 12161219. doi: 10.1126/science.1231097

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Imai, S., and Guarente, L. (2010). Ten years of NAD-dependent SIR2 family
deacetylases: implications for metabolic diseases. Trends Pharmacol. Sci. 31,
212220. doi: 10.1016/j.tips.2010.02.003

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Jeong, H., Cohen, D. E., Cui, L., Supinski, A., Savas, J. N., Mazzulli, J. R., et al.
(2011). Sirt1 mediates neuroprotection from mutant huntingtin by activation of
the TORC1 and CREB transcriptional pathway. Nat. Med. 18, 159165. doi:
10.1038/nm.2559

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Jiang, J. C., Jaruga, E., Repnevskaya, M. V., and Jazwinski, S. M. (2000). An

intervention resembling caloric restriction prolongs life span and retards aging in
yeast. FASEB J. 14, 21352137. doi: 10.1096/fj.00-0242fje

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Jiang, M., Wang, J., Fu, J., Du, L., Jeong, H., West, T., et al. (2011). Neuroprotective
role of Sirt1 in mammalian models of Huntingtons disease through activation of
multiple Sirt1 targets. Nat. Med. 18, 153158. doi: 10.1038/nm.2558

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Jin, L., Galonek, H., Israelian, K., Choy, W., Morrison, M., Xia, Y., et al. (2009).
Biochemical characterization, localization, and tissue distribution of the longer
form of mouse SIRT3. Protein Sci. 18, 514525. doi: 10.1002/pro.50

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Julien, C., Tremblay, C., Emond, V., Lebbadi, M., Salem, N. Jr., Bennett, D. A., et al.
(2009). Sirtuin 1 reduction parallels the accumulation of tau in Alzheimer
disease. J. Neuropathol. Exp. Neurol. 68, 4858. doi:
10.1097/NEN.0b013e3181922348

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Jung, C. H., Ro, S. H., Cao, J., Otto, N. M., and Kim, D. H. (2010). mTOR regulation
of autophagy. FEBS Lett. 584, 12871295. doi: 10.1016/j.febslet.2010.01.017

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Kaeberlein, M., McVey, M., and Guarente, L. (1999). The SIR2/3/4 complex and
SIR2 alone promote longevity in Saccharomyces cerevisiae by two different
mechanisms. Genes Dev. 13, 25702580. doi: 10.1101/gad.13.19.2570

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Kakefuda, K., Fujita, Y., Oyagi, A., Hyakkoku, K., Kojima, T., Umemura, K., et al.
(2009). Sirtuin 1 overexpression mice show a reference memory deficit, but not
neuroprotection. Biochem. Biophys. Res. Commun. 387, 784788. doi:
10.1016/j.bbrc.2009.07.119

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Kendrick, A. A., Choudhury, M., Rahman, S. M., McCurdy, C. E., Friederich, M., Van
Hove, J. L., et al. (2011). Fatty liver is associated with reduced SIRT3 activity and
mitochondrial protein hyperacetylation. Biochem. J. 433, 505514. doi:
10.1042/BJ20100791

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Kim, D., Nguyen, M. D., Dobbin, M. M., Fischer, A., Sananbenesi, F., Rodgers, J. T.,
et al. (2007). SIRT1 deacetylase protects against neurodegeneration in models
for Alzheimers disease and amyotrophic lateral sclerosis. EMBO J. 26, 3169
3179. doi: 10.1038/sj.emboj.7601758

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Kim, S. C., Sprung, R., Chen, Y., Xu, Y., Ball, H., Pei, J., et al. (2006). Substrate and
functional diversity of lysine acetylation revealed by a proteomics survey. Mol.
Cell. 23, 607618. doi: 10.1016/j.molcel.2006.06.026

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Klar, A. J., and Fogel, S. (1979). Activation of mating type genes by transposition
in Saccharomyces cerevisiae. Proc. Natl. Acad. Sci. U.S.A. 76, 45394543. doi:
10.1073/pnas.76.9.4539

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Koubova, J., and Guarente, L. (2003). How does calorie restriction work? Genes
Dev. 17, 313321. doi: 10.1101/gad.1052903

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Lan, F., Cacicedo, J. M., Ruderman, N., and Ido, Y. (2008). SIRT1 modulation of the
acetylation status, cytosolic localization, and activity of LKB1. Possible role in
AMP-activated protein kinase activation. J. Biol. Chem. 283, 2762827635. doi:
10.1074/jbc.M805711200

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Lanza, I. R., Short, D. K., Short, K. R., Raghavakaimal, S., Basu, R., Joyner, M. J., et
al. (2008). Endurance exercise as a countermeasure for aging. Diabetes 57,
29332942. doi: 10.2337/db08-0349

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Lassmann, H. (2010). Axonal and neuronal pathology in multiple sclerosis: what

have we learnt from animal models. Exp. Neurol. 225, 28. doi:
10.1016/j.expneurol.2009.10.009

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Lavu, S., Boss, O., Elliott, P. J., and Lambert, P. D. (2008). Sirtuins novel
therapeutic targets to treat age-associated diseases. Nat. Rev. Drug Discov. 7,
841853. doi: 10.1038/nrd2665

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Law, I. K., Liu, L., Xu, A., Lam, K. S., Vanhoutte, P. M., Che, C. M., et al. (2009).
Identification and characterization of proteins interacting with SIRT1 and SIRT3:
implications in the anti-aging and metabolic effects of sirtuins. Proteomics 9,
24442456. doi: 10.1002/pmic.200800738

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Lee, I. H., Cao, L., Mostoslavsky, R., Lombard, D. B., Liu, J., Bruns, N. E., et al.
(2008). A role for the NAD-dependent deacetylase Sirt1 in the regulation of
autophagy. Proc. Natl. Acad. Sci. U.S.A. 105, 33743379. doi:
10.1073/pnas.0712145105

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Lescai, F., Blanche, H., Nebel, A., Beekman, M., Sahbatou, M., Flachsbart, F., et al.
(2009). Human longevity and 11p15.5: a study in 1321 centenarians. Eur. J. Hum.
Genet. 17, 15151519. doi: 10.1038/ejhg.2009.54

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Liu, D., Gharavi, R., Pitta, M., Gleichmann, M., and Mattson, M. P. (2009).
Nicotinamide prevents NAD+ depletion and protects neurons against
excitotoxicity and cerebral ischemia: NAD+ consumption by SIRT1 may endanger
energetically compromised neurons. Neuromol. Med. 11, 2842. doi:
10.1007/s12017-009-8058-1

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Lombard, D. B., Alt, F. W., Cheng, H. L., Bunkenborg, J., Streeper, R. S.,
Mostoslavsky, R., et al. (2007). Mammalian Sir2 homolog SIRT3 regulates global
mitochondrial lysine acetylation. Mol. Cell. Biol. 27, 88078814. doi:
10.1128/MCB.01636-07

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Luo, J., Nikolaev, A. Y., Imai, S., Chen, D., Su, F., Shiloh, A., et al. (2001). Negative
control of p53 by Sir2alpha promotes cell survival under stress. Cell 107, 137
148. doi: 10.1016/S0092-8674(01)00524-4

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Madeo, F., Tavernarakis, N., and Kroemer, G. (2010). Can autophagy promote
longevity? Nat. Cell Biol. 12, 842846. doi: 10.1038/ncb0910-842

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Markert, C. D., Kim, E., Gifondorwa, D. J., Childers, M. K., and Milligan, C. E.
(2010). A single-dose resveratrol treatment in a mouse model of amyotrophic
lateral sclerosis. J. Med. Food 13, 10811085. doi: 10.1089/jmf.2009.0243

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Masoro, E. J. (2000). Caloric restriction and aging: an update. Exp. Gerontol. 35,
299305. doi: 10.1016/S0531-5565(00)00084-X

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Mihaylova, M. M., and Shaw, R. J. (2011). The AMPK signalling pathway

coordinates cell growth, autophagy and metabolism. Nat. Cell Biol. 13, 1016
1023. doi: 10.1038/ncb2329

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Min, S. W., Cho, S. H., Zhou, Y., Schroeder, S., Haroutunian, V., Seeley, W. W., et
al. (2010). Acetylation of tau inhibits its degradation and contributes to
tauopathy. Neuron 67, 953966. doi: 10.1016/j.neuron.2010.08.044

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Morris, K. C., Lin, H. W., Thompson, J. W., and Perez-Pinzon, M. A. (2011).

Pathways for ischemic cytoprotection: role of sirtuins in caloric restriction,
resveratrol, and ischemic preconditioning. J. Cereb. Blood Flow Metab. 31, 1003
1019. doi: 10.1038/jcbfm.2010.229

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Navarro, A., and Boveris, A. (2007). The mitochondrial energy transduction

system and the aging process. Am. J. Physiol. Cell Physiol. 292, C670C686. doi:
10.1152/ajpcell.00213.2006

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Nemoto, S., Fergusson, M. M., and Finkel, T. (2005). SIRT1 functionally interacts
with the metabolic regulator and transcriptional coactivator PGC-1{alpha}. J.
Biol. Chem. 280, 1645616460. doi: 10.1074/jbc.M501485200

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Nisoli, E., Tonello, C., Cardile, A., Cozzi, V., Bracale, R., Tedesco, L., et al. (2005).
Calorie restriction promotes mitochondrial biogenesis by inducing the expression
of eNOS. Science 310, 314317. doi: 10.1126/science.1117728

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Okawara, M., Katsuki, H., Kurimoto, E., Shibata, H., Kume, T., and Akaike, A.
(2007). Resveratrol protects dopaminergic neurons in midbrain slice culture from
multiple insults. Biochem. Pharmacol. 73, 550560. doi:
10.1016/j.bcp.2006.11.003

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Onyango, P., Celic, I., McCaffery, J. M., Boeke, J. D., and Feinberg, A. P. (2002).
SIRT3, a human SIR2 homologue, is an NAD-dependent deacetylase localized to
mitochondria. Proc. Natl. Acad. Sci. U.S.A. 99, 1365313658. doi:
10.1073/pnas.222538099

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Palacios, O. M., Carmona, J. J., Michan, S., Chen, K. Y., Manabe, Y., Ward, J. L. III, et
al. (2009). Diet and exercise signals regulate SIRT3 and activate AMPK and PGC-
1alpha in skeletal muscle. Aging (Albany NY) 1, 771783.

Pubmed Abstract | Pubmed Full Text

Pallas, M., Pizarro, J. G., Gutierrez-Cuesta, J., Crespo-Biel, N., Alvira, D., Tajes, M.,
et al. (2008). Modulation of SIRT1 expression in different neurodegenerative
models and human pathologies. Neuroscience 154, 13881397. doi:
10.1016/j.neuroscience.2008.04.065

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Pallos, J., Bodai, L., Lukacsovich, T., Purcell, J. M., Steffan, J. S., Thompson, L. M.,
et al. (2008). Inhibition of specific HDACs and sirtuins suppresses pathogenesis in
a Drosophila model of Huntingtons disease. Hum. Mol. Genet. 17, 37673775.
doi: 10.1093/hmg/ddn273

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Parker, J. A., Arango, M., Abderrahmane, S., Lambert, E., Tourette, C., Catoire, H.,
et al. (2005). Resveratrol rescues mutant polyglutamine cytotoxicity in nematode
and mammalian neurons. Nat. Genet. 37, 349350. doi: 10.1038/ng1534

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Patel, N. V., Gordon, M. N., Connor, K. E., Good, R. A., Engelman, R. W., Mason, J.,
et al. (2005). Caloric restriction attenuates Abeta-deposition in Alzheimer
transgenic models. Neurobiol. Aging. 26, 9951000. doi:
10.1016/j.neurobiolaging.2004.09.014

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Perry, V. H., Lunn, E. R., Brown, M. C., Cahusac, S., and Gordon, S. (1990).
Evidence that the Rate of Wallerian degeneration is controlled by a single
autosomal dominant gene. Eur. J. Neurosci. 2, 408413. doi: 10.1111/j.1460-
9568.1990.tb00433.x

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Petersen, K. F., Befroy, D., Dufour, S., Dziura, J., Ariyan, C., Rothman, D. L., et al.
(2003). Mitochondrial dysfunction in the elderly: possible role in insulin
resistance. Science 300, 11401142. doi: 10.1126/science.1082889

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Picard, F., Kurtev, M., Chung, N., Topark-Ngarm, A., Senawong, T., Machado De
Oliveira, R., et al. (2004). Sirt1 promotes fat mobilization in white adipocytes by
repressing PPAR-gamma. Nature 429, 771776. doi: 10.1038/nature02583

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Puigserver, P., Wu, Z., Park, C. W., Graves, R., Wright, M., and Spiegelman, B. M.
(1998). A cold-inducible coactivator of nuclear receptors linked to adaptive
thermogenesis. Cell 92, 829839. doi: 10.1016/S0092-8674(00)81410-5

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Qin, W., Chachich, M., Lane, M., Roth, G., Bryant, M., de Cabo, R., et al. (2006a).
Calorie restriction attenuates Alzheimers disease type brain amyloidosis in
Squirrel monkeys (Saimiri sciureus). J. Alzheimers Dis. 10, 417422.

Pubmed Abstract | Pubmed Full Text

Qin, W., Yang, T., Ho, L., Zhao, Z., Wang, J., Chen, L., et al. (2006b). Neuronal
SIRT1 activation as a novel mechanism underlying the prevention of Alzheimer
disease amyloid neuropathology by calorie restriction. J. Biol. Chem. 281, 21745
21754. doi: 10.1074/jbc.M602909200

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Qiu, X., Brown, K., Hirschey, M. D., Verdin, E., and Chen, D. (2010). Calorie
restriction reduces oxidative stress by SIRT3-mediated SOD2 activation. Cell
Metab. 12, 662667. doi: 10.1016/j.cmet.2010.11.015

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Ramadori, G., Lee, C. E., Bookout, A. L., Lee, S., Williams, K. W., Anderson, J., et
al. (2008). Brain SIRT1: anatomical distribution and regulation by energy
availability. J. Neurosci. 28, 99899996. doi: 10.1523/JNEUROSCI.3257-08.2008

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Raval, A. P., Dave, K. R., and Perez-Pinzon, M. A. (2006). Resveratrol mimics

ischemic preconditioning in the brain. J. Cereb. Blood Flow Metab. 26, 1141
1147. doi: 10.1038/sj.jcbfm.9600262

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Rodgers, J. T., Lerin, C., Haas, W., Gygi, S. P., Spiegelman, B. M., and Puigserver, P.
(2005). Nutrient control of glucose homeostasis through a complex of PGC-
1alpha and SIRT1. Nature 434, 113118. doi: 10.1038/nature03354

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Rogina, B., and Helfand, S. L. (2004). Sir2 mediates longevity in the fly through a
pathway related to calorie restriction. Proc. Natl. Acad. Sci. U.S.A. 101, 15998
16003. doi: 10.1073/pnas.0404184101

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Rose, G., Dato, S., Altomare, K., Bellizzi, D., Garasto, S., Greco, V., et al. (2003).
Variability of the SIRT3 gene, human silent information regulator Sir2 homologue,
and survivorship in the elderly. Exp. Gerontol. 38, 10651070. doi:
10.1016/S0531-5565(03)00209-2

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Rosen, D. R. (1993). Mutations in Cu/Zn superoxide dismutase gene are
associated with familial amyotrophic lateral sclerosis. Nature 364, 362. doi:
10.1038/364362c0

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Sakakibara, I., Fujino, T., Ishii, M., Tanaka, T., Shimosawa, T., Miura, S., et al.
(2009). Fasting-induced hypothermia and reduced energy production in mice
lacking acetyl-CoA synthetase 2. Cell Metab. 9, 191202. doi:
10.1016/j.cmet.2008.12.008

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Salminen, A., and Kaarniranta, K. (2012). AMP-activated protein kinase (AMPK)

controls the aging process via an integrated signaling network. Ageing Res.
Rev. 11, 230241. doi: 10.1016/j.arr.2011.12.005

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Salminen, A., Kauppinen, A., Suuronen, T., and Kaarniranta, K. (2008a). SIRT1
longevity factor suppresses NF-kappaB-driven immune responses: regulation of
aging via NF-kappaB acetylation? Bioessays 30, 939942. doi:
10.1002/bies.20799

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Salminen, A., Ojala, J., Huuskonen, J., Kauppinen, A., Suuronen, T., and
Kaarniranta, K. (2008b). Interaction of aging-associated signaling cascades:
inhibition of NF-kappaB signaling by longevity factors FoxOs and SIRT1. Cell. Mol.
Life Sci. 65, 10491058. doi: 10.1007/s00018-008-7461-3

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Sasaki, Y., Vohra, B. P., Baloh, R. H., and Milbrandt, J. (2009). Transgenic mice
expressing the Nmnat1 protein manifest robust delay in axonal degeneration in
vivo. J. Neurosci. 29, 65266534. doi: 10.1523/JNEUROSCI.1429-09.2009

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Satoh, A., Brace, C. S., Ben-Josef, G., West, T., Wozniak, D. F., Holtzman, D. M., et
al. (2010). SIRT1 promotes the central adaptive response to diet restriction
through activation of the dorsomedial and lateral nuclei of the hypothalamus. J.
Neurosci. 30, 1022010232. doi: 10.1523/JNEUROSCI.1385-10.2010

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Sauve, A. A. (2009). Pharmaceutical strategies for activating sirtuins. Curr.

Pharm. Des. 15, 4556. doi: 10.2174/138161209787185797

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Sauve, A. A., Wolberger, C., Schramm, V. L., and Boeke, J. D. (2006). The
biochemistry of sirtuins. Annu. Rev. Biochem. 75, 435465. doi:
10.1146/annurev.biochem.74.082803.133500

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Schlicker, C., Gertz, M., Papatheodorou, P., Kachholz, B., Becker, C. F., and
Steegborn, C. (2008). Substrates and regulation mechanisms for the human
mitochondrial sirtuins Sirt3 and Sirt5. J. Mol. Biol. 382, 790801. doi:
10.1016/j.jmb.2008.07.048

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Schmitz, M. L., Mattioli, I., Buss, H., and Kracht, M. (2004). NF-kappaB: a
multifaceted transcription factor regulated at several levels. Chembiochem 5,
13481358. doi: 10.1002/cbic.200400144

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Schwer, B., Bunkenborg, J., Verdin, R. O., Andersen, J. S., and Verdin, E. (2006).
Reversible lysine acetylation controls the activity of the mitochondrial enzyme
acetyl-CoA synthetase 2. Proc. Natl. Acad. Sci. U.S.A. 103, 1022410229. doi:
10.1073/pnas.0603968103

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Schwer, B., Eckersdorff, M., Li, Y., Silva, J. C., Fermin, D., Kurtev, M. V., et al.
(2009). Calorie restriction alters mitochondrial protein acetylation. Aging Cell 8,
604606. doi: 10.1111/j.1474-9726.2009.00503.x

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Schwer, B., North, B. J., Frye, R. A., Ott, M., and Verdin, E. (2002). The human
silent information regulator (Sir)2 homologue hSIRT3 is a mitochondrial
nicotinamide adenine dinucleotide-dependent deacetylase. J. Cell Biol. 158, 647
657. doi: 10.1083/jcb.200205057

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Shi, T., Wang, F., Stieren, E., and Tong, Q. (2005). SIRT3, a mitochondrial sirtuin
deacetylase, regulates mitochondrial function and thermogenesis in brown
adipocytes. J. Biol. Chem. 280, 1356013567. doi: 10.1074/jbc.M414670200

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Shimazu, T., Hirschey, M. D., Huang, J. Y., Ho, L. T., and Verdin, E. (2010). Acetate
metabolism and aging: an emerging connection. Mech. Ageing Dev. 131, 511
516. doi: 10.1016/j.mad.2010.05.001

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Shimokawa, I., and Trindade, L. S. (2010). Dietary restriction and aging in
rodents: a current view on its molecular mechanisms. Aging Dis. 1, 89107.

Pubmed Abstract | Pubmed Full Text

Shindler, K. S., Ventura, E., Dutt, M., Elliott, P., Fitzgerald, D. C., and Rostami, A.
(2010). Oral resveratrol reduces neuronal damage in a model of multiple
sclerosis. J. Neuroophthalmol. 30, 328339. doi:
10.1097/WNO.0b013e3181f7f833

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Shindler, K. S., Ventura, E., Rex, T. S., Elliott, P., and Rostami, A. (2007). SIRT1
activation confers neuroprotection in experimental optic neuritis. Invest.
Ophthalmol. Vis. Sci. 48, 36023609. doi: 10.1167/iovs.07-0131

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Shulga, N., Wilson-Smith, R., and Pastorino, J. G. (2010). Sirtuin-3 deacetylation

of cyclophilin D induces dissociation of hexokinase II from the mitochondria. J.
Cell Sci. 123, 894902. doi: 10.1242/jcs.061846

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Sinclair, D. A. (2002). Paradigms and pitfalls of yeast longevity research. Mech.

Ageing Dev. 123, 857867. doi: 10.1016/S0047-6374(02)00023-4

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Singh, K. K. (2004). Mitochondrial dysfunction is a common phenotype in aging

and cancer. Ann. N. Y. Acad. Sci. 1019, 260264. doi: 10.1196/annals.1297.043

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Someya, S., Yu, W., Hallows, W. C., Xu, J., Vann, J. M., Leeuwenburgh, C., et al.
(2010). Sirt3 mediates reduction of oxidative damage and prevention of age-
related hearing loss under caloric restriction. Cell 143, 802812. doi:
10.1016/j.cell.2010.10.002

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Swerdlow, R. H. (2007). Treating neurodegeneration by modifying mitochondria:

potential solutions to a complex problem. Antioxid. Redox Signal. 9, 1591
1603. doi: 10.1089/ars.2007.1676

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Swerdlow, R. H. (2011). Brain aging, Alzheimers disease, and

mitochondria. Biochim. Biophys. Acta 1812, 16301639. doi:
10.1016/j.bbadis.2011.08.012

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Tang, B. L. (2009). Sirt1s complex roles in neuroprotection. Cell. Mol.
Neurobiol. 29, 10931103. doi: 10.1007/s10571-009-9414-2

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Tao, R., Coleman, M. C., Pennington, J. D., Ozden, O., Park, S. H., Jiang, H., et al.
(2010). Sirt3-mediated deacetylation of evolutionarily conserved lysine 122
regulates MnSOD activity in response to stress. Mol. Cell. 40, 893904. doi:
10.1016/j.molcel.2010.12.013

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Tissenbaum, H. A., and Guarente, L. (2001). Increased dosage of a sir-2 gene

extends lifespan in Caenorhabditis elegans. Nature 410, 227230. doi:
10.1038/35065638

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Viswanathan, M., and Guarente, L. (2011). Regulation of Caenorhabditis

elegans lifespan by sir-2.1 transgenes. Nature 477, E1E2. doi:
10.1038/nature10440

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Wakeling, L. A., Ions, L. J., and Ford, D. (2009). Could Sirt1-mediated epigenetic
effects contribute to the longevity response to dietary restriction and be
mimicked by other dietary interventions? Age (Dordr.) 31, 327341. doi:
10.1007/s11357-009-9104-5

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Wang, J., Fivecoat, H., Ho, L., Pan, Y., Ling, E., and Pasinetti, G. M. (2010). The role
of Sirt1: at the crossroad between promotion of longevity and protection against
Alzheimers disease neuropathology. Biochim. Biophys. Acta 1804, 16901694.
doi: 10.1016/j.bbapap.2009.11.015

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Wang, J., Ho, L., Qin, W., Rocher, A. B., Seror, I., Humala, N., et al. (2005a).
Caloric restriction attenuates beta-amyloid neuropathology in a mouse model of
Alzheimers disease. FASEB J. 19, 659661. doi: 10.1096/fj.04-3182fje

Pubmed Abstract | Pubmed Full Text

Wang, J., Zhai, Q., Chen, Y., Lin, E., Gu, W., McBurney, M. W., et al. (2005b). A
local mechanism mediates NAD-dependent protection of axon degeneration. J.
Cell Biol. 170, 349355. doi: 10.1083/jcb.200504028

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Wang, P., Xu, T. Y., Guan, Y. F., Tian, W. W., Viollet, B., Rui, Y. C., et al. (2011).
Nicotinamide phosphoribosyltransferase protects against ischemic stroke through
SIRT1-dependent adenosine monophosphate-activated kinase pathway. Ann.
Neurol. 69, 360374. doi: 10.1002/ana.22236

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Wang, S., Xing, Z., Vosler, P. S., Yin, H., Li, W., Zhang, F., et al. (2008). Cellular
NAD replenishment confers marked neuroprotection against ischemic cell death:
role of enhanced DNA repair. Stroke 39, 25872595. doi:
10.1161/STROKEAHA.107.509158

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Wareski, P., Vaarmann, A., Choubey, V., Safiulina, D., Liiv, J., Kuum, M., et al.
(2009). PGC-1{alpha} and PGC-1{beta} regulate mitochondrial density in
neurons. J. Biol. Chem. 284, 2137921385. doi: 10.1074/jbc.M109.018911

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Wood, J. G., Rogina, B., Lavu, S., Howitz, K., Helfand, S. L., Tatar, M., et al. (2004).
Sirtuin activators mimic caloric restriction and delay ageing in
metazoans. Nature 430, 686689. doi: 10.1038/nature02789

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Xiong, S., Salazar, G., Patrushev, N., and Alexander, R. W. (2011). FoxO1
mediates an autofeedback loop regulating SIRT1 expression. J. Biol. Chem. 286,
52895299. doi: 10.1074/jbc.M110.163667

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Yang, Y., Hubbard, B. P., Sinclair, D. A., and Tong, Q. (2010). Characterization of
murine SIRT3 transcript variants and corresponding protein products. J. Cell.
Biochem. 111, 10511058. doi: 10.1002/jcb.22795

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Yang, Y. H., Chen, Y. H., Zhang, C. Y., Nimmakayalu, M. A., Ward, D. C., and
Weissman, S. (2000). Cloning and characterization of two mouse genes with
homology to the yeast Sir2 gene. Genomics 69, 355369. doi:
10.1006/geno.2000.6360

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text

Yeung, F., Hoberg, J. E., Ramsey, C. S., Keller, M. D., Jones, D. R., and Frye, R. A.,
et al. (2004). Modulation of NF-kappaB-dependent transcription and cell survival
by the SIRT1 deacetylase. EMBO J. 23, 23692380. doi:
10.1038/sj.emboj.7600244

Pubmed Abstract | Pubmed Full Text | CrossRef Full Text