ABSTRAK

STUDI MEKANISME KERJA SENYAWA

ALFA MANGOSTIN DAN XANTON
SEBAGAI ANTIDIABETES

Oleh
Welly Ratwita
NIM: 30712302
(Program Studi Doktor Farmasi)

Tujuan: Penelitian ini bertujuan untuk mengetahui mekanisme kerja α-mangostin

dan xanton sebagai antidiabetes. Diuraikan efek α-mangostin dan xanton terhadap
kadar glukosa darah puasa dan insulin plasma, regenerasi Pulau Langerhans,
Glucose Transporter (GLUT)-4, serta Peroxixome proliferator-activated receptor
gamma (PPAR-γ).
Metode: Uji toleransi glukosa dilakukan menggunakan tikus jantan galur Wistar
yang dibagi secara acak menjadi 9 kelompok uji (n=5 ekor/kelompok), yaitu
kelompok normal, kontrol (diberi D-glukosa 2 g/kgbb), kelompok perlakuan (diberi
α-mangostin atau xanton dengan dosis 5, 10 atau 20 mg/kgbb), kelompok
pembanding diberi glibenklamid 0,45 mg/kgbb, 30 menit sebelum diberi D-
glukosa 2 g/kgbb. Perubahan kadar glukosa darah diamati pada menit ke-60 hingga
ke-150. Seluruh kelompok diberi pakan standar dan air minum sesuai kebutuhan.
Uji toleransi insulin menggunakan tikus jantan galur Wistar, yang dibagi secara
acak menjadi 9 kelompok uji (n=5 ekor/kelompok), yaitu kelompok normal,
kontrol, kelompok perlakuan (diberi α-mangostin atau xanton dengan dosis 5, 10
atau 20 mg/kgbb), kelompok pembanding diberi metformin 4,5 mg/kgbb.
Kelompok normal hanya mendapat pakan standar dan air minum sesuai kebutuhan.
Kelompok kontrol, perlakuan dan pembanding diberi diet emulsi tinggi lemak 1
ml/hari, selama 10 hari, kemudian pada hari ke-11 diberi insulin 0,05 IU/kgbb
intraperitoneal. Kadar glukosa darah pada tikus diukur enam kali setelah pemberian
insulin, setiap 30 menit selama 150 menit. Uji kadar glukosa darah puasa, kadar
insulin dan regenerasi Pulau Langerhans menggunakan mencit Swiss Webster
jantan, yang dibagi menjadi 10 kelompok secara acak, yaitu kelompok normal,
kontrol (hanya diinduksi aloksan monohidrat 70 mg/kgbb), kelompok perlakuan
diberi berbagai dosis α-mangostin atau xanton (dengan dosis 5,10, atau 20
mg/kgbb) selama 21 hari. Kadar glukosa darah puasa dan kadar insulin plasma
diperiksa pada hari ke-21, jaringan pankreas diambil dan disimpan dalam dapar
formalin 10%, dilanjutkan dengan pewarnaan Gomori. Jantung juga diambil dan
disimpan dalam dapar formalin 10%, kemudian diwarnai dengan proses
imunohistokimia, untuk mengamati ekspresi GLUT-4. Kultur sel adiposit
dilakukan untuk mengamati uji ekspresi GLUT-4 dan PPAR-γ. Studi in vitro
dilakukan untuk mengukur inhibisi α-glukosidase, dan glikasi. Absorbansi diukur
menggunakan spektrofotometer pada panjang gelombang 530 nm.
Hasil: Kadar glukosa darah di setiap kelompok perlakuan pada menit ke-90 hingga
ke-150 menurun secara signifikan ketika dibandingkan dengan kelompok kontrol
(p <0,05). Hal ini menunjukkan bahwa terjadi peningkatan toleransi glukosa pada
setiap kelompok yang diberi perlakuan. Koefisien tes toleransi insulin (KTTI) pada
semua kelompok perlakuan berbeda secara signifikan (p <0,05) jika
dibandingkan dengan kelompok kontrol, kecuali xanton 5 mg/kgbb. Hal ini
menunjukkan bahwa α-mangostin 5, 10 dan 20 mg/kgbb, xanton 10 dan 20
mg/kgbb, serta metformin, meningkatkan toleransi insulin pada tikus putih yang
diberikan diet emulsi lemak 10 hari. Efek α-mangostin dan xanton (5, 10 dan 20
mg/kgbb) pada kadar glukosa darah puasa tidak berbeda secara signifikan dari
glibenklamid dan metformin. Hasil uji kadar insulin plasma menunjukkan bahwa
terdapat perbedaan yang signifikan antara α-mangostin 20 mg/kgbb jika
dibandingkan dengan kontrol. Uji regenerasi Pulau Langerhans menunjukkan
perbedaan yang signifikan antara α-mangostin dan xanton (5, 10 dan 20 mg/kgbb)
dengan kelompok kontrol. Ekspresi GLUT-4 pada sel-sel otot jantung tikus yang
diberi dengan α-mangostin (5, 10, dan 20 mg/kgbb) atau xanton (10, dan 20
mg/kgbb) meningkat secara signifikan bila dibandingkan dengan kelompok
kontrol, dan tidak berbeda secara signifikan dari glibenklamid dan metformin.
Ekspresi GLUT-4 pada adiposit yang mendapat 3,13 mM; 6,25 mM atau 25 mM α-
mangostin juga meningkat setara dengan pioglitazon. Ekspresi PPAR-γ pada sel
adiposit yang mendapat α-mangostin dan xanton meningkat jika dibandingkan
kontrol. Ekspresi PPARγ pada sel adiposit kelompok yang mendapat α-mangostin
7,28 mM lebih baik daripada pioglitazon. Nilai IC50 α-mangostin terhadap α-
glukosidase lebih baik daripada akarbosa dan xanton, sedangkan nilai IC50
piridoksamin terhadap reaksi glikasi lebih baik daripada α-mangostin dan xanton.
Kesimpulan: Alfa mangostin dan xanton adalah dua senyawa yang dapat
meningkatkan toleransi glukosa dan insulin. Efek antidiabetes tampak pada
turunnya kadar glukosa darah puasa, meningkatnya kadar insulin plasma dan
regenerasi Pulau Langerhans, ekspresi GLUT-4 pada otot jantung dan adiposit,
ekspresi PPAR-γ pada sel adiposit, dan juga sebagai inhibitor α-glukosidase dan
glikasi.

Kata kunci: α-mangostin, xanton, toleransi glukosa, toleransi insulin, kadar

glukosa darah puasa, kadar insulin plasma, regenerasi Pulau Langerhans,
GLUT-4, PPAR-γ, α-glukosidase, glikasi.
 ABSTRACT

STUDY OF THE MECHANISM OF ACTION

OF ALPHA MANGOSTIN AND XANTHONE COMPUNDS
AS ANTIDIABETIC

By
Welly Ratwita
NIM 30712302
(Doctoral Study of Pharmacy)

Objective: This research elaborated the role of α-mangostin and xanthone on

glucose and insulin tolerance, fasting blood glucose, plasma insulin and islet of
Langerhans regeneration in alloxan induced diabetic mice, Glucose Transporter
(GLUT)-4 by measuring GLUT-4 expression in cardiac muscle cell and adipocyte
culture and peroxisome proliferator–activated receptor (PPAR)-γ expression on
adipocyte culture. We also elaborated the effect of α-mangostin and xanthone as
inhibitor for -glucosidase and glication, in order to find -mangostin and
xanthone mechanism of action as antidiabetic agent.
Methods: Glucose tolerance test were conducted using male wistar rat divided into
9 groups, which were normal, control (D-Glucose induced only), glibenclamide at
0.45 m/kgbw, α mangostin and xanthone (at different dose ofdoses 5, 10, 20
mg/kgbw). All groups were induced with D-glucose 2 g/kgbw orally. Thirty minutes
later, blood glucose level changes were observed at 60th to 150th minute. The insulin
tolerance test used 45 male Wistar rats, which were divided randomly into the
normal group, control group received only high fatty emulsion diet, treatment
group which received high fat emulsion diet and α-mangostin or xanthone at 5, 10
or 20 mg/kgbw, or metformin at dose of 45 mg/kgbw. Normal group received
only regular drinking water, while control and treatment groups received a high fat
emulsion for 10 days. Blood sugar level were measured six times after insulin
administration (0.05 IU/kgbw) intraperitoneally, every 30 minutes for 150 minutes.
Fasting blood glucose, insulin and islet of Langerhans examination were conducted
using male Swiss webster mice, divided into 10 groups randomly, which were
normal, control (alloxan induced only), glibenclamide at 0.65 mg/kgbw, various
doses of -mangostin and xanthone (at 5, 10, or 20 mg/kgbw) for 21
days, then they were sacrificed by cervical decapitation on day 21st. Fasting blood
glucose and insulin plasma were then measured. Pancreatic tissues were isolated
from sacrificed animals, and then fixed in neutral buffered formalin. Histologic
observations of the Islet of Langerhans were performed after staining using Gomori
staining method. The heart was also isolated and kept in formaline buffered to be
subsequently stained using immunohistochemistry to observed GLUT-4 expression.
Adypocite was cultured to observed GLUT-4 and PPAR-γ expression. In vitro study
was carried out to measure inhibition of α-glucosidase and glycation using
spectrophotometer 530 nm.
Results: Normal group (non diabetic) responded slightly to the administration of
glucose in glucose tolerance test. Blood glucose level in every group in the 90th to
150th minute decreased significantly when compared to the control group (p
<0.05). This showed that glucose tolerance increased in all treated groups
althought they were treated with high glucose consentration. Coefficient of insulin
tolerance test in α-mangostin groups were significantly different (p<0.05) when
compared to the control group, except in group treated with xanthone at dose 5
mg/kgbw. This suggests that α-mangostin 5, 10 and 20 mg/kgbw, xanthone 10 and
20 mg/kgbw, as well as metformin at 45 mg/kgbw, increased insulin tolerance in
Wistar rats given fatty emulsion for ten days. The effects of α-mangostin and
xanthone on fasting blood glucose were not significantly different from
glibenclamide and metformin. Plasma insulin analysis showed that there were no
significant difference between 20 mg/kgbw α-mangostin compared with control.
Alpha mangostin, xanthone had magnificant effect on the islet of Langerhans
compared to control group and glibenclamid and metformin groups. GLUT-4
expressions in mice cardiac muscle cells treated with α-mangostin, xanthone,
glibenclamide, and metformin significantly increased when compared to the control
group, except in group treated with xanthone at dose 5 mg/kgbw. These increasing
were not significantly different from glibenclamide and metformin. GLUT-4
expressions also increased in adipocytes treated with 3.13 mM; 6.25 mM and 25
mM α-mangostin. All treatment group results were significantly different when
compared with control. The effect of α-mangostin on GLUT-4 expression was better
than that of xanthone. Almost similar as pioglitazone, α-mangostin and xanthone
increased PPAR-γ expression in adipocyte, but the effect of xanthone was not as
good as that of α-mangostin or pioglitazone effect. IC50 of α-mangostin for α-
glucosidase was better than acarbose and xanthone, while IC50 of pyridoxamine for
glycation was better than α-mangostin and xanthone.
Conclusion: Alpha mangostin and xanthone are two substances that may improve
glucose and insulin tolerance. In addition they showed antidiabetic effect by
improving fasting blood glucose level, insulin plasma and the regeneration of islet
of Langerhans, GLUT-4 expression in cardiac muscle and adipocyte, PPAR-γ
expression in adipocyte, and also α-glucosidase and glycation inhibiting activities.

Keywords: α-mangostin, xanthone, glucose tolerance, insulin tolerance, fasting

blood glucose, plasma insulin, the islet of Langerhans, GLUT-4, PPAR-γ,
α-glucosidase, glycation.