Efek Prokteksi Coenzim Q10 pada Methamphetamine-Induksi

Apoptosis pada Tikus Dewasa Jantan

Jurnal Reading

DISUSUN OLEH:
Davien Utoyo (133307010099)

PEMBIMBING:
dr. Adhanyani Lubis, Sp. KJ

Kepaniteraan Klinik Senior (KKS) Bagian Ilmu Kesehatan Jiwa

Fakultas Kedokteran Prima Indonesia
Rumah Sakit Royal Prima
Medan
2018
 Novelty in Biomedicine

Original Article

Protective Effect of Coenzyme Q10 on Methamphetamine-Induced

Apoptosis in Adult Male Rats

Fatemeh Gholipour1, Jamal Shams*1, Alireza Zahiroddin1

1
Imam Hossein hospital, Behavioral Science Research Center of Shahid Beheshti University of Medical Science (SBUMS), Tehran, Iran

Received: 20 December, 2016; Accepted: 07 May, 2017

Abstract
Background: The negative consequence of methamphetamine abuse is due to neuropathologic changes in the brain,
which reduces dopaminergic neurons and result in damage to different brain areas. Neurotoxicity induced by
methamphetamine increases the oxidative stress and associated with neuronal apoptosis. The role of the antioxidant
coenzyme Q10 probably produces its neuroprotective effects. Therefore, the purpose of the present study was to
examine the protective effect of coenzyme Q10 on methamphetamine-induced apoptosis in adult male rats.
Materials and Methods: Fifty Wistar eight-week adult rats randomly divided into 5 groups: Healthy control,
methamphetamine injection (Meth), methamphetamine injection and CoQ10 5mg/kg treatment (Meth+Post CoQ10
5mg/kg), methamphetamine injection and CoQ10 10mg/kg treatment (Meth+Post CoQ10 10mg/kg),
methamphetamine injection and CoQ10 20mg/kg treatment (Meth+Post CoQ10 20mg/kg). Methamphetamine with a
purity of 96% with a dosage of 20 mg/kg was injected Intraperitoneal. Coenzyme Q10 for three treatment groups was
injected intraperitoneally for 14 days in a dosage of 5, 10 and 20 mg/kg/day. The protein expressions of Baxand Bcl2
were evaluated by western blotting technique.
Results: Bax protein expression was significantly lower in Meth+Post CoQ10 5mg/kg (p=0.010) and so Meth+Post
CoQ10 10mg/kg (p=0.004) comparing to Meth group. In addition, Bcl2 protein expression was significantly higher
in Meth+Post CoQ10 5mg/kg comparing to Meth group (p=0.018). However, there were no significant differences
between control and CoQ10 treatment groups. Bax/Bcl2 ratio was significantly lower in Meth+Post CoQ10 5mg/kg
(p=0.005), Meth+Post CoQ10 10mg/kg (p=0.008) and Meth+Post CoQ10 20mg/kg (p=0.044) comparing to Meth
group.
Conclusion: We suggest that CoQ10 reduces the methamphetamine-induced apoptosis in the striatum of the rats
through the reduction of apoptotic factors and increase of anti-apoptotic pathways.
Keywords: Addiction, Apoptosis, Coenzyme Q10, Methamphetamine

*Corresponding Author: Jamal Shams, MD. Associate professor of Psychiatry, Behavioral Research Center and department of psychiatry,
Shahid Beheshti University of Medical Science (SBUMS). Email: Tehran, Iran. j_shams@yahoo.com

Please cite this article as: Gholipour F, Shams J, Zahiroddin A. Protective Effect of Coenzyme Q10 on Methamphetamine-Induced Apoptosis
in Adult Male Rats. Novel Biomed. 2017;5(3):127-32.

Introduction have strong stimulant effects on CNS. Its popularity is

Addiction and substance abuse have negative effects on
human’s life, including physical and mental health, and
social, legal, and emotional status1. Methamphetamine is
an addictive psychoactive drug which has been proven to
NBM
increasing in US and other parts of the world because of
low price2. Methamphetamine abuse is becoming a
public health problem in global level and it is estimated
that there are 15-16 million methamphetamine abuser
around the world, which is making it the second abused
127 Novelty in Biomedicine 2017, 3, 127-32
Gholipour et al. Protective Effect of Coenzyme Q10 on Methamphetamine-Induced Apoptosis …
substance3. Euphoria caused by methamphetamine is
accompanied by loss of appetite and body temperature,
paranoia, aggression, and increased sense of joy4. In
humans, methamphetamine abuse has cognitive and
neurologic damages and cardiovascular complications5.
Methamphetamine can pass through blood-brain barrier
and is probably able to damage dopaminergic neurons.
Methamphetamine injection can cause prolonged release
of dopamine in humans and animals6.
Oxidative stress, mitochondrial disruption, apoptosis,
neural inflammation, and toxicity mediated by NMDA
receptors are mechanisms generally mentioned in
neurotoxicity of dopaminergic neurons7. Chronic abuse
of methamphetamine causes decrease in dopamine
transporters in cortex, caudate and putamen, and decrease
in serotonin transporters8, as well as microglial activation
in different areas of brain9 and morphologic changes like
a decrease in gray matter and hypertrophy of white
matter10. In many studies there are convincing evidences
in support of methamphetamine damage to different areas
of brain and neuron loss10-12. Evidences suggest that
methamphetamine neurotoxicity involves reactive
oxygen species (ROS), reactive nitrogen species (RNS)
and lower-hand oxidative stress mechanisms activation13.
Methamphetamine enters dopaminergic neurons via
dopamine transporters and shifts dopamine vesicle.
Moved amines can be oxided in enzyme and non-enzyme
forms and makes reacting Quoinons and ROS and
increase oxidative stress14. Thus, ROS causes death of
neurons due to damage to cellular parts including DNA,
RNA, and proteins. Also mitochondrial disruption in
neural destruction caused by methamphetamine has been
noticed15. Methamphetamine is lipophilic cation
molecule that can infiltrate into mitochondria and stays
there. In fact, methamphetamine reduces mitochondrial
membrane potential and level of I, III, and VI complex of
electron transport chain in primary human cells16 which
is followed by decreased ATP in brain17. Pro apoptotic
proteins like Bax, Bad and Bid are increased and anti-
apoptotic proteins like Bcl-2 and Bcl-XL are decreased
after injection of methamphetamine17. Plus,
methamphetamine not only causes cellular death via
apoptosis, but also via necrotic mechanisms18.
Studies suggest that medication modalities in order to
prevent and treat destructive effects of methamphetamine
need attention to pathways that form substrates of
methamphetamine-induced toxicity. Co-enzyme Q10 is
necessary enzyme in the electron transport chain and an
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effective agent for neuroprotection in neurodegenerative
diseases which increases plasma levels of Q10 co-
enzyme and prevents decrease in dopamine when added
to diet19. In recent years use of co-enzyme Q10 has been
in center of attention for treatment of neurodegenerative
diseases.co-enzyme Q10 probably acts as a
neuroprotective agent by its anti-oxidant and free radical
neutrizing effects. Plus that, co-enzyme Q10 is an
electron receptor in complex I of electron transport
chain20. Co-enzyme Q10 protects neurons from oxidative
stress and prevents reduction in membrane potential of
mitochondrial membrane21. Studies have shown that co-
enzyme Q10 reduces neural damage of hippocampus in
regions CA1, CA2, and CA3. Clinical studies also
showed that co-enzyme Q10 can protect dopaminergic
system of striatum and slows progression of disability in
Parkinson disease22. Since apoptosis caused by
methamphetamine-induces neurotoxicity can damage
various parts of brain, especially dopaminergic part, and
there are findings on protective effects of co-enzyme Q10
on neurologic system, in this survey, we study the
protective effect of co-enzyme Q10 on apoptosis caused
by injection on methamphetamine on adult male rat.
Methods
This is an experimental-interventional study on heads of
50 adult male 8-weeks old wistar rats, weighing 200±20
gr. Rats were housed in standard cages and controlled
conditions of light (12 hr darkness, 12 hr light),
temperature 22±3℃ and humidity around 45%, with free
access to food and water. All steps were done by
supervision of Ethics committee of Iran University of
Medical Sciences and according to the moral protocol of
experiments on lab animals. Samples are divided
randomly into these five groups: Control,
methamphetamine injection (meth), methamphetamine
injection + post 5mg/kg co-enzyme Q10,
methamphetamine injection + post 10mg/kg co-enzyme
Q10, and methamphetamine injection + post 20mg/kg co-
enzyme Q10.
Methamphetamine was obtained from police and had
96% purity. It was solved in physiologic serum and was
injected totally 20 mg/kg within 2 days (10 mg each day,
5 mg 2 times a day with 12hr intervals)
intraperitoneally3. For verification of injection, mobility
test was performed. When injected, if animal passed 10
times around a 30cm basket, an injection would be
128 Novelty in Biomedicine 2017, 3, 127-32
Protective Effect of Coenzyme Q10 on Methamphetamine-Induced Apoptosis … Gholipour et al.

considered effective. Then the Co-enzyme Q10 injection enzyme Q10 group (p=0.018) was significantly higher
as a post-treatment was starting after one day from the
last injection of Methamphetamine. Co-enzyme Q10
provided from Sigma Co (St. louis, MO, USA) and
solved with concentration of 100mg/cc in sesame oil. The
solution of Co-enzyme Q10 was injected within 14 days
in 5, 10, 20 mg/kg/day intraperitoneally for three post-
treatment groups. The control group just received
physiologic serum without Methamphetamine but the
same volume to Meth group.
For the purpose of studying of expression of Bax and
Bcl2 proteins in striatal region, the animals was
sacrificed for provide brain tissue. All of the animals
anesthetized by intraperitoneal injection of ketamine and
xylazine in 8:1 proportion and after cutting their head and
removing their brain, striatal part was dissected and Figure 1. Expression of Bax protein in between groups,* significant
washed by normal saline immediately and put striatal
parts in microtubes and freeze in liquid nitrogen and then
into a refrigerator with -80℃ and stored until western
blotting them. Process had to done in less than 4 minutes
after cutting head for each rat. For studying expression of
Bax Bcl2 proteins, it was first derived and after that,
using western blotting technique, protein bands removed
from SDS-PAGE gel via electrophoresis were moved to a
nitrocellulose membrane and in next step, were detected
using primary and secondary antibodies23. For
Quantitation of protein bands TotalLab Version 1.10
software was used. Data analysis was done using SPSS
software. All data are reported with medium ± standard
deviation. Kolmogrof-smirnof test was done to evaluate
normal distribution of data and if distribution was
normal, then parametric tests were used. Difference
between groups was studied using unilateral variance
analysis test and Tukey. Dunnett`s T3post hoc tests was
done to compare the groups. The level of significance
was set at p ≤ 0.05.

Results
As shown in figure 1, expression of Bax protein in
control group (p=0.012), meth +post 5mg/kg (p=0.010),
meth + post 10mg/kg (p=0.004) was significantly less
than Meth group. But there is no significant difference
between meth+ post 20mg/kg and Meth (p=0.056). There
is also no significant difference between groups which
were treated with co-enzyme Q10 and control group
(p>0.05).
Expression of Bcl2 protein in meth + post 5mg/kg co- Figure 2. Expression of BCL2 protein in between groups,*

NBM 129 Novelty in Biomedicine 2017, 3, 127-32

Gholipour et al. Protective Effect of Coenzyme Q10 on Methamphetamine-Induced Apoptosis …
than meth group, but there was no significant difference
between groups treated by co-enzyme Q10 and control
group (p>0.05, figure 2).
Ratio of Bax/Bcl2 in the control group (p=0.014), meth
+post 5mg/kg co-enzyme Q10 (p= 0.005), meth + post
10mg/kg co-enzyme Q10 (p=0.008), meth + post
20mg/kg co-enzyme Q10 (p=0.044) was significantly
lower than Meth group, but there was no significant
difference between groups treated by co-enzyme Q10 and
control group (p>0.05).
Discussion
Deep stimulation of the brain is another modality for
treatment of addiction and various parts of brain
including nucleus accumbens, subthalamic nucleus,
dorsal striatum, medial prefrontal cortex, and other parts
of the brain are targeted24. How the methamphetamine
increased apoptosis induction in striatum? There are
convincing evidences that negative psycho-neural
consequences of methamphetamine abuse is partly
because of pathologic neurological changes in brain of
abusers25. Dopamine release caused by
methamphetamine is oxidized by monoamine oxidase
and dihydroxyphenil acetic acid and hydrogen peroxide
are produced26. Hydrogen peroxide interacts with metal
ions and produces highly toxic hydroxyl free radical via
phenton reaction. Thus, reacting oxygen species (ROS)
including hydroxyl free radicals cause neuron death via
damage to cellular parts like DNA, RNA and proteins.
Methamphetamine is a lipophilic cation molecule which
can diffuse into mitochondria and stays there.
Methamphetamine also decreases the activity of complex
2 and 4 of electron transport chain swiftly which is
accompanied by a decrease of ATP in brain17. In this
study methamphetamine injection caused apoptosis in
striatum which was consistent with previous studies27.
This apoptosis might be because of various factors like
oxidants, oxygen radicals and nitric oxide, which are
originated from excessive release of glutamate and
dopamine28. It is shown that methamphetamine causes an
increase in pro apoptotic proteins like Bax, Bad and Bid,
but decreases anti apoptotic proteins like Bcl-2 and Bcl-
Xl17. In this study, compatible with previous surveys, we
demonstrated that methamphetamine induces apoptotic
factors like Bax in rats. We also showed that induction of
these apoptotic factors is reduced by co-enzyme Q10
anti-oxidant. So there is probability that apoptosis of
dopaminergic neurons in striatum of rats is reduced. How
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Co-enzyme Q10 reduced apoptosis induction? Co-
enzyme Q10 is a necessary enzyme in the electron
transport chain and an effective neuroprotective agent in
neurodegenerative disease. Adding co-enzyme Q10 to the
diet increases its plasma level and prevents dopamine
decrease19. It also acts as important antioxidants in
mitochondria and lipid membranes. There is a
remarkable interest in using co-enzyme Q10 in
neurodegenerative diseases.co-enzyme Q10 protects
neurons from oxidative stress and prevents reduction in
mitochondrial membrane potential21. In animal models of
Parkinson disease has protective function29. Co-enzyme
Q10 also protects dopaminergic neurons from
mitochondrial membrane depolarization and rotenone-
induced death30. A mechanism of co-enzyme Q10
neuroprotective effects is acting as an uncoupling
proteins co-factor31.
Co-enzyme Q10 suppresses apoptosis caused by
oxidative stress and mitochondrial permeability32. It also
stops Bax connection with mitochondria and cytochrome
C release from mitochondria33. Co-enzyme Q10
significantly reduces indices of lipid peroxidation in
plasma, erythrocytes, liver and brain of rats34. So there is
probability that in this study, co-enzyme Q10
supplementation protects against methamphetamine-
induced dopaminergic apoptosis by increasing anti-
oxidant activity and neutralizing free radicals. It has also
has been shown that co-enzyme Q10 reduces cocaine and
methamphetamine induced neurotoxicity23.
In this study, we demonstrated induction of apoptosis in
striatum after acute injections of methamphetamine,
because levels of Bax protein in Meth group were
increased. Although treatment with doses of 5 and 10 of
co-enzyme Q10 after injection of methamphetamine
caused a decrease in Bax protein levels in comparison
with Meth group. Probably protective effects of co-
enzyme Q10 after injection of methamphetamine is due
to its anti-apoptotic mechanisms. In addition, to
mentioned apoptotic factors, higher levels of Bcl-2
protein in treatment with dose 5 of co-enzyme Q10 after
injection of methamphetamine was observed. Also ratio
of Bax/Bcl2 was significantly lower in groups of
treatment with dose 5, 10 and 20 of co-enzyme Q10 in
comparison with Meth group which is a reflection of an
increase in anti-apoptotic status.
130 Novelty in Biomedicine 2017, 3, 127-32
Protective Effect of Coenzyme Q10 on Methamphetamine-Induced Apoptosis …
Conclusion
Overall, the findings of this study suggest that co-enzyme
Q10 reduces apoptosis induced by methamphetamine
injection in striatum of rats via reduction in apoptotic
factors like Bax and increase in anti-apoptotic pathways.
We noticed that co-enzyme Q10 reduces apoptosis
induction after injection of methamphetamine in male
rats. We hope that treatment with co-enzyme Q10 after
addiction to methamphetamine be helpful in addicted
patients.
Acknowledgment
This survey has been done by the financial support of
Behavioral Science Research Center Shahid Beheshti
University of medical sciences.
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132 Novelty in Biomedicine 2017, 3, 127-32
 Efek Prokteksi Coenzim Q10 pada Methamphetamine-Induksi Apoptosis pada

Tikus Dewasa Jantan

Fatemeh Gholipour1, Jamal Shams*1, Alireza Zahiroddin1

1
Rumah Sakit Imam Hossein, Pusat Penelitian Ilmu Perilaku di Shahid Beheshti Ilmu

Medis (SBUMS), Tehran Iran

Menerima: 20 Desember, 2016; Diterima: 07 Mei, 2017

Abstrak

Latar Belakang: Konsenkuensi negatif pada penyalahgunaan methamphetamine

dapat disebabkan oleh perubahan neuropatologik pada otak, dimana berkurangnya

neuron dopaminergik dan menghasilkan kerusakan otak di berbagai area yang berbeda.

Neurotoksisitas diinduksi oleh peningkatan stress oksidatif pada methamphetamine

dan dikaitkan dengan apoptosis neuronal. Peran aksi oksidan pada co-enzim Q10

mungkin memproduksi efek neuroproktektif. Oleh karena itu, tujuan dari penelitian ini

untuk menilai efek perlindungan pada co-enzim Q10 pada methamphetamine-induksi

apoptosis pada tikus jantan dewasa.

Material dan Metode: Lima puluh Wistar delapan minggu tikus dewasa secara acak

terbagi atas 5 kelompok: Kontrol Sehat, injeksi methamphetamine (Meth), injeksi

methamphetamine dan CoQ10 5mg/kg pengobatan (Meth+Post CoQ10 10mg/kg),

injeksi methamphetamine dan C0Q10 20mg/kg pengobatan (Meth+Post CoQ10

20mg/kg). Methamphetamine dengan kemurnian 96% pada dosis 20mg/kg disuntikan

di intraperitoneal. Co-enzim Q10 untuk tiga kelompok pengobatan dapat disuntikan di

intraperitoneal untuk 14 hari pada dosis 5, 10, dan 20mg/kg/hari. Ekspresi protein pada

Baxand Bcl2 dapat di evaluasi dengan teknik western blotting.

Hasil: Ekspresi Bax protein signifikan rendah pada Meth+Post CoQ10 5mg/kg

(p=0.010) dan Meth+Post C0Q10 10mg/kg (p=0.004) dibandingkan dengan kelompok

Meth. Penambahan, ekspresi protein Bcl2 signifikan tinggi pada Meth+Post CoQ10

5mg/kg dibandingkan dengan kelompok Meth (p=0.018). Namun, tidak ada perbedaan

yang signifikan antara kontrol dan kelompok pengobatan CoQ10. Rasio Bac/Bcl2

signifikan rendah pada Meth+Post CoQ10 5mg/kg (p=0.005), Meth+Post CoQ10

10mg/kg (p=0.008) dan Meth+Post CoQ10 20 mg/kg (p=0.044) dibandingkan dengan

kelompok Meth.

Kesimpulan: Menurut kami CoQ10 mengurangi methamphetamine-induksi apoptosis

di striatum pada tikus melalui pengurangan faktor apoptosis dan peningkatan jalur anti-

apoptotis.

Kata Kunci: Kecanduan, Apoptosis, Coenzim Q10, Methamphetamine

Pendahuluan

Kecanduan dan penyalahgunaan substansi dapat menimbukan efek negatif pada

kehidupan manusia, termasuk fisik dan kesehatan mental, dan sosial, hukum, dan status

emosi. (Golmirzaei J, Amiri S, et all. 2014). Methamphetamine adalah obat adiktif

psikoaktif dimana dapat mencegah efek stimulasi yang kuat pada CNS. Ini sangat

popular meningkat pada US dan bagian dunia yang lain karena harga yang rendah.

(United Nations Office on Drugs and Crime. 2005). Penyalahgunaan

methamphetamine menjadi masalah kesehatan publik pada tingkat global dan

diperkirakan 15-16 juta penyalahgunaan methamphetamine disekitar dunia, yang

membuat penyalahgunaan substansi methamphetamine dalam hitungan detik.

(Krasnova IN, Cadet JL. 2009). Euphoria penyebab dari methamphetamine yang

dikaitkan dengan hilangnya nafsu makan dan temperatur tubuh, paranoia, agresi, dan

peningkatan rasa senang. (Homer BD, Solomon TM, et all. 2008). Pada manusia,

penyalahgunaan methamphetamine mempunyai kognitif dan kerusakan neurologik dan

komplikasi kardiovaskular. (Darke S, Kaye Set, et all. 2008). Methamphetamine dapat

melewati pertahanan sawar otak (Blood Brain Barrier) dan mungkin dapat merusak

neuron dopaminergik. Injeksi methamphetamine dapat menyebabkan pelepasan

dopamin yang lama pada manusia dan hewan. (Kita T, Wagner GC, et all. 2003).

Stress oksidatif, gangguan mitokondria, apoptosis, inflamasi neural, dan toksisitas yang

dimediasi oleh reseptor NMDA sebagai mekanisme umum sebagai neurotoksisitas

pada neuron dopaminergik. (Riddle EL, Fleckenstein AE. 2006). Penyalahgunaan

methamphetamine yang lama dapat menyebabkan menurunnya transpor dopamin di

korteks, caudate dan putamen, dan penurunan transpor serotonin. (McCann UD,
Kuwabara H, et all. 2008), seperti pengaktifan mikroglia pada area otak yang berbeda.

(Sekine Y, Ouchi Y, Sugihara G, et all. 2008) dan perubahan morfologi seperti

penurunan gray matter dan hipertropi pada white matter. (Thompson PM, Hayashi

KM, et all. 2004). Banyak penelitian yang membuktikan bahwa methamphetamine

menyebabkan kerusakaan pada berbagai area di otak dan hilangnya neuron (Thompson

PM, Hayashi KM, et all. 2004) (Kuczenski R, Everall IP, et all. 2007) (Chou J, Luo Y,

et all. 2008). Bukti menunjukkan bahwa neurotoksisitas methamphetamine melibatkan

aktivasi kembali spesis oksigen (ROS), spesis aktivasi nitrogen (RNS) dan pengaktifan

mekanisme oksidatif stess pada tangan rendah. (Stephans SE, Yamamoto BK, et all.

1994). Methamphetamine masuk ke neuron dopaminergik melalui jalur dopamin

transport dan pengantian vesikel dopamin. Perpindahan amina dapat oksidasi enzim

dan non-enzim yang terbentuk dan membuat Quinon beraksi kembali dan ROS dan

peningkatan oksidatif stress (Michel PP, Hefti F. 1990). Demikian, penyebab kematian

ROS pada neuron menyebabkan kerusakan bagian selular termasuk DNA, RNA, dan

protein. Juga gangguan mitokondria pada dekstrusi saraf yang mengakibatkan

methamphetamine sudah diumumkan (Wu CW, Ping YH, et all 2007).

Methamphetamine adalah molekul kation lipofilik dapat infiltrat ke dalam mitokondria

dan menetap. Faktanya, methamphetamine mengurangi membran mitokondria yang

potensial dan tingkat I, III, dan kompleks VI pada rantai transport pada sel primer

manusia (Potula R, Hawkins BJ, et all. 2010) yang diikuti dengan adanya penurunan

ATP dalam otak. (Oliveira MT, Rego AC, et all. 2002). Pro apoptosis protein seperti

Bax, Bad dan Bid meningkat dan anti-apoptosis protein seperti Bcl-2 dan Bcl-XL

menurun setelah injeksi methamphetamine. (Oliveira MT, Rego AC, et all. 2002).
Ditambah, methamphetamine tidak hanya penyebab kematian selular melalui jalur

apoptosis, tetapi juga melalui mekanisme nekrosis (Gold MS, Kobeissy FH, et all.

2009)

Peneliti menyimpulkan bahwa modalitas pengobatan untuk mencegah dan mengobati

efek destruktif pada methamphetamine yang memerlukan perhatian pada jalur yang

membentuk substrat pada methamphetamine-induksi toksisitas. Co-enzim Q10 adalah

enzim penting dalam rantai transport elektron dan agen efektif untuk neuroproteksi

pada penyakit neurodegeneratif dimana peningkatan plasma pada Q10 co-enzim dan

mencegah penurunan dopamin ditambah dengan diet (Turunen M, Olsson J, et all.

2004). Dalam beberapa tahun terakhir co-enzim Q10 telah digunakan sebagai pusat

perhatian untuk pengobatan pada penyakit neurodegeneratif. Co-enzim Q10 mungkin

berperan sebagai agen neuroproteksi dengan anti-oksidan dan efek netral radikal bebas.

Ditambah, co-enzim Q10 adalah reseptor elektron kompleks I pada rantai transport

elektron (Thrash B, Karuppagounder SS, et all. 2010). Pelindung neuron co-enzim Q10

dari oksidasi stress dan mencegah penurunan potensial membran pada membran

mitokondria (Somayajulu M, McCarthy S, et all. 2005). Peneliti menunjukkan co-

enzim Q10 mengurangi kerusakan neural pada hippocampus di bagian CA1, CA2, dan

CA3. Studi klinis juga menunjukkan co-enzim Q10 dapat melindungi sistem

dopaminergik pada striatum dan memperlambat proses pada disabilitas penyakit

Parkinson (Shults CW. 2005). Ketika apoptosis penyebab methamphetamine-induksi

neurotoksisitas dapat merusak beberapa bagian otak, khususnya bagian dopaminergik,

dan penemuan efek proteksi pada co-enzim Q10 pada sistem neurologik, pada survei
ini, kita belajar efek proteksi pada co-enzim Q10 pada apoptosis disebabkan oleh

injeksi methamphetamine pada tikus jantan dewasa.

Metode

Experimen ini-studi intervensional pada 50 tikus wistar jantan dewasa usia 8 minggu,

berat 200±20 gr. Tikus rumah dalam sangkar standar dan kondisi kontrol ringan (12

jam gelap, 12 jam terang) temperatur 22±30C dan kelembaban sekitar 45% dengan

akses bebas pada makanan dan minuman. Semua langkah telah dilakukan pengawasan

oleh supervisi pada Komite Etik Universitas Iran bagian Ilmu Medis dan menurut

protokol moral pada eksperimen di laboratorium binatang. Sampel dibagi secara acak

ke dalam 5 kelompok: Kontrol, injeksi methamphetamine (meth), injeksi

methamphetamine + post 5mg/kg co-enzim Q10, injeksi methamphetamine + post

10mg/kg co-enzim Q10, dan injeksi methamphetamine + post 20mg/kg co-enzim Q10.

Methamphetamine diperoleh dari kepolisian dan memiliki 96% kemurnian. Itu sudah

dipecahkan kedalam serum fisiologis dan jumlah injeksi 20mg/kg dalam 2 hari (10 mg

tiap hari, 5 mg 2 kali dalam satu hari dengan interval 12 jam) secara intraperitoneal

(Krasnova IN, Cadet JL. 2009). Untuk verifikasi injeksi, tes mobilitas ditunjukkan.

Saat disuntikkan, jika hewan melewati 10 kali sekitar keranjang 30cm, suntikan akan

dianggap efektif. Kemudian suntikan co-enzim Q10 sebagai pasca rawatan sebagai

awal setelah satu hari dari injeksi methamphetamine terakhir. Co-enzim Q10

disediakan dari Sigma Co (St. louis, MO, USA) dan konsentrasi yang dipecahkan

100mg/CC dalam sesame oil. Solusio Co-enzim Q10 disuntikan sekitar 14 hari dalam

5, 10, 20 mg/kg/hari di intraperitoneal untuk tiga kelompok perlakuan. Kelompok

kontrol baru menerima serum fisiologis tanpa methamphetamine tetapi volume yang

sama untuk kelompok Meth.

Untuk tujuan belajar pada ekspresi Bax dan protein Bcl2 pada bagian striatal, hewan

yang dikorbankan untuk jaringan otak. Semua hewan di anastesi dengan injeksi

intraperitoneal pada ketamin dan xylazine dengan proporsi 8:1 dan setelah memotong

kepala mereka dan mengeluarkan otak mereka, bagian striatal dibedah dan dicuci

dengan normal salin segera mungkin dan meletakan bagian striatal dalam mikrotube

dan beku dalam cairan nitrogen dan kemudian kedalam kulkas dengan -800C dan

disimpan sampai mereka di western blotting. Proses harus dilakukan dengan waktu

yang kurang dari 4 menit setelah memotong masing-masing kepala tikus. Untuk

pembelajaran ekspresi protein Bax Bcl2, itu pertama diturunkan dan setelah itu,

menggunakan teknik western blotting, ikatan protein dilepaskan dari gel SDS-PAGE

melalui elektrophoresis dipindahkan ke membran nitroselulosa dan langkah kemudian,

di deteksi menggunakan antibodi primer dan sekunder (Klongpanichapak S,

Govitrapong P, et all. 2006). Untuk kuantitas perangkat lunak ikatan protein TotalLab

Versi 1.10 yang digunakan. Analisis data dapat dikerjakan dengan perangkat lunak

SPSS. Semua data telah dilaporkan dengan medium ± standar deviasi. Kolmogrof-

smirnof tes dilakukan untuk evaluasi distribusi data normal dan jika distribusi normal,

kemudian tes parametrik digunakan. Perbedaan antara kelompok peneliti

menggunakan tes analisis variasi unilateral dan Tukey. Tes Dunnett’s T3 post hoc dapat

dilakukan untuk membandingkan kelompok. Tingkat signifikansi ditetapkan pada

p≤0.05.
Hasil

Seperti yang terlihat pada figur 1, ekspresi pada protein Bax dalam kelompok kontrol

(p=0.012), meth + post 5mg/kg (p=0.010), meth + post 10mg/kg (p=0.004) kurang

signifikan dari kelompok Meth. Tetapi tidak ada perbedaan signifikan diantara meth +

post 20mg/kg dan Meth (p=0.056). Tidak ada perbedaan yang signifikan juga diantara

kelompok yang diobati dengan co-enzim Q10 dan grup kontrol (p>0.05).

Ekspresi pada Bcl2 protein dalam meth + post 5mg/kg co-enzim kelompok Q10

(p=0.018) signifikan tinggi dari kelompok meth, tetapi tidak ada perbedaan signifikan

antara kelompok yang diobati dengan co-enzim Q10 dan kelompok kontrol (p>0.05,

figur 2).

Rasio Bax/Bcl2 dalam kelompok kontrol (p=0.014), meth + post 5mg/kg co-enzim Q10

(p=0.005), meth + post 10mg/kg co-enzim (p=0.008), meth + post 20mg/kg co-enzim

Q10 (p=0.044) signifikan rendah dari kelompok Meth, tetapi tidak ada perbedaan

kelompok yang signifikan antara kelompok yang diobati dengan co-enzim Q10 dan

kelompok kontrol (p>0.05).

Figur 1. Ekspresi protein Bax antara kelompok,* signifikan

Figur 2. Ekspresi protein BCL2 antar kelompok,*

Diskusi

Stimulasi otak dalam adalah modalitas untuk pengobatan kecanduan dan berbagai

bagian pada otak termasuk nukleus accumbens, subthalamic nukleus, dorsal striatum,

korteks medial prefrontal, dan target otak bagian lain (Salamone JD. 1992). Bagaimana

methamphetamine menginduksi peningkatan induksi apoptosis di striatum? Ada bukti

yang meyakinkan konsekuensi psiko-neural negatif pada sebagian penyalahgunaan

methamphetamine dapat menyebabkan perubahan neurologis patologik dalam otak

pada penyalahguna (Scott JC, Woods SP, et all. 2007). Pelepasan dopamin

menyebabkan oksidasi methamphetamine oleh oksidasi monoamin dan asam

dihidroksipenil asetik dan hidrogen peroksida terproduksi (Olanow CW, Tatton WG.

1999). Hidrogen peroksida berinteraksi dengan ion metal dan produksi radikal bebas

yang tinggi toksik melalui reaksi phenton. Demikian, reaksi oksigen spesies (ROS)

termasuk radikal bebas hidroksi menyebabkan kematian neuron melalui kerusakan

bagian selular seperti DNA, RNA dan protein. Methamphetamine adalah molekul

kation lipopilik yang dapat difusi kedalam mitokondria dan menetap.

Methamphetamine juga dapat menurun pada aktivitas kompleks 2 dan 4 rantai transpor

elektron dengan cepat yang didampingi oleh penurunan ATP dalam otak (Oliveira MT,

Rego AC, et all. 2002). Dalam penelitian ini injeksi methamphetamine disebabkan oleh

apoptosis dalam striatum yang konsisten dengan peneliti sebelumnya (Deng X, Wang

Y, et all. 2001). Apoptosis ini dapat disebabkan oleh variasi faktor seperti oksidan,

radikal oksigen dan nitrit oksida, yang mengatur dari pelepasan glutamat dan dopamin

yang berlebihan (Cadet JL, Krasnova IN. 2009). Ini menunjukkan penyebab

methamphetamine adalah peningkatan protein pro apoptosis seperti Bax, Bad dan Bid,
tetapi penurunan protein anti apoptotis seperti Bcl-2 dan Bcl-XI (Oliveira MT, Rego

AC, et all. 2002). Penelitian ini, sesuai dengan survei sebelumnya, kami menunjukkan

bahwa apoptotis induksi methamphetamine faktor seperti Bax pada tikus. Kamu juga

menunjukkan bahwa induksi pada faktor apoptotis mengurangi anti oksidan co-enzim

Q10. Jadi ada kemungkinan apoptosis pada neuron dopaminergik pada striatum tikus

yang berkurang. Bagaimana Co-enzim Q10 berkurang induksi apoptosis? Co-enzim

Q10 adalah enzim penting dalam rantai transpor elektron dan agen neuroproteksi yang

efektif pada penyakit neurodegeneratif. Penambahan co-enzim Q10 untuk asupan

meningkat pada tingkat plasma dan mencegah dopamin berkurang (Turunen M, Olsson

J, et all. 2004). Itu juga bertindak sebagai antioksidan yang penting dalam mitokondria

dan membran lipid. Ada ketertarikan saat menggunakan co-enzim Q10 dalam penyakit

neurodegeneratif. Co-enzim Q10 melindungi neuron dari oksidasi stress dan mencegah

reduksi membran potensial mitokondria (Somayajulu M, McCarthy S, et all. 2005).

Contoh hewan pada penyakit parkinson mempunyai fungsi prokteksi (Ebadi M,

Brown-Borg H, et all. 2005). Co-enzim Q10 juga melindungi neuron dopaminergik

dari depolarisasi membran mitokondria dan kematian rotenone-induksi (Moon Y, Lee

KH, et all. Moon Y, Lee KH, 2005). Mekanisme co-enzim Q10 efek neuroprokteksi

bertindak sebagai melepaskan co-faktor protein (Echtay KS, Winkler E, et all. 2000).

Supresi co-enzim Q10 disebabkan oleh stress oksidatif dan permeabilitas mitokondria

(Alleva R, Tomasetti M, et all. 2001). Itu juga berhenti hubungan Bax dengan

mitokondria dan pelepasan sitokrom C dari mitokondria (Naderi J, Somayajulu-Nitu

M, et all. 2006). Co-enzim Q10 signifikan menunjukkan berkurangnya peroksidasi

lipid dalam plasma, eritrosit, liver, dan otak tikus (Tomasetti M, Alleva R, et all. 2001).
Jadi ada kemungkinan bahwa penelitian ini, suplementasi co-enzim Q10 melindungi

kembali induksi methamphetamine apoptosis dopaminergik dengan meningkatkan

aktivitas antioksidan dan menetralisir radikal bebas. Itu juga ditunjukkan co-enzim Q10

berkurang kokain dan induksi methamphetamine neurotoksisitas (Klongpanichapak S,

Govitrapong P, et all. 2006).

Dalam penelitian ini, kami menunjukkan induksi apoptosis dalam striatum setelah

injeksi methamphetamine akut, karena tingkat protein Bax pada kelompok Meth

meningkat. Meskipun pengobatan dengan dosis dan pada co-enzim setelah injeksi

methamphetamine menyebabkan penurunan tingkat Bax protein dibandingkan dengan

kelompok Meth. Mungkin efek proteksi pada co-enzim Q10 setelah injeksi

methamphetamine dikarenakan mekanisme anti-apoptotis, tingkat tinggi pada protein

Bcl-2 pada pengobatan dengan dosis dengan co-enzim Q10 setelah injeksi

methamphetamine menyebabkan mekanisme anti-apoptotis. Selain itu, untuk faktor

apoptosis, Bcl-2 protein tingkat tinggi dengan pengobatan co-enzim Q10 dosis setelah

injeksi methamphetamine diamati. Juga pada rasio Bax/Bcl2 signifikan rendah pada

kelompok pengobatan dengan dosis 5, 10 dan 20 pada co-enzim Q10 dibandingkan

dengan kelompok Meth yang merupakan refleksi pada peningkatan status anti-

apoptosis.

Kesimpulan

Secara keseluruhan, temuan penelitian ini menyarankan bahwa berkurangnya apoptosis

pada co-enzim Q10 induksi oleh injeksi methamphetamine di striatum tikus melalui

reduksi pada apoptosis faktor seperti Bax dan peningkatan jalur anti-apoptosis. Kami

memberitahukan bahwa co-enzim Q10 mengurangi induksi apoptosis setelah injeksi

methamphetamin pada tikus jantan. Kami berharap pengobatan dengan co-enzim Q10

setelah kecanduan methamphetamine bisa membantu pada pasien yang kecanduan.

Pernyataan Resmi

Survei ini telah selesai dengan dukungan finansial dari Pusat Penelitian Ilmu Perilaku

Universitas Sains Shahid Beheshti