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Sains Malaysiana 47(11)(2018): 2799–2809 http://dx.doi.org/10.17576/jsm-2018-4711-23
The Effect of Tocotrienol-Rich Fraction
on the Expression of Glutathione S-Transferase Isoenzymes in Mice
Liver (Kesan Pecahan Kaya Tokotrienol kepada
Ekspresi Isoenzim Glutation S-Transferase di dalam Hepar Tikus) AHMED ATIA1,2,
NADIA
SALEM
ALRAWAIQ1
& AZMAN ABDULLAH1* 1Department of Pharmacology,
Faculty of Medicine, Universiti Kebangsaan Malaysia, Jalan Yaacob
Latif, Bandar Tun Razak, 56000 Cheras, Kuala Lumpur, Federal Territory,
Malaysia 2Department of Anesthesia
and Intensive Care, Faculty of Medical Technology, Tripoli University,
Tripoli, Libya Diserahkan: 29
Mac 2018/Diterima: 30 Julai 2018 ABSTRACT Glutathione S-transferase isoenzymes
(GSTs) catalyze the conjugation reaction between glutathione
and electrophilic compounds. GSTs are involved in the detoxification
of toxic and carcinogenic compounds, thus protecting the body
from toxic injuries. Tocotrienols are part of the vitamin E family
and is believed to possess potent antioxidant activity. The objective
of this study was to determine the effect of increasing doses
of tocotrienol rich fraction (TRF)
supplementation on liver GSTs gene and protein expression. A total
of 30 male ICR white mice were divided into five
groups (n=6 for each group) and given treatment for 14 days through oral
supplementation. Groups were divided as follows: - three groups
administered with TRF at doses of 200, 500 and 1000 mg/kg
respectively, a positive control group administered with 100 mg/kg,
butylated hydroxyanisole (BHA) and a control group administered
with only the vehicle (corn oil). At day 15, the mice were sacrificed
and their livers isolated. Total RNA was extracted from the liver and
quantitative real-time polymerase chain reaction (qPCR) assays
were performed to analyze GSTs gene expression. Total liver protein was also extracted
and the protein expression of GSTs was determined by Western
blotting. The results showed that TRF oral supplementation caused
a significant dose-dependent increase in liver GST isoenzymes
gene and protein expression, compared to controls. In conclusion,
TRF
oral supplementation for 14 days resulted in increased
gene and protein expression of GST isoenzymes in mice liver dose-dependently, with the
highest expression seen in mice treated with 1000 mg/kg TRF. Keywords: Glutathione S-transferase;
isoenzymes; liver; tocotrienols; TRF ABSTRAK Glutation S-transferase (GST)
isoenzim merupakan enzim yang memangkinkan tindak balas konjugasi
antara glutation dan sebatian hidrofilik. GST terlibat dalam detoksifikasi
sebatian toksik dan karsinogenik dan oleh itu melindungi badan
daripada kecederaan toksik. Tokotrienol adalah sebahagian daripada
keluarga besar vitamin E dan dipercayai mempunyai aktiviti antioksidan
yang poten. Objektif kajian ini adalah untuk menentukan kesan
pemberian dos secara meningkat fraksi kaya tokotrienol (TRF) terhadap pengekspresan gen dan protein
beberapa isoenzim GST di hepar. Sebanyak 30 ekor mencit
putih jantan ICR dibahagikan kepada lima kumpulan (n=6
untuk setiap kumpulan) dan dirawat secara suplementasi oral selama
14 hari. Kumpulan dibahagikan seperti berikut: - tiga kumpulan
dirawat dengan pelbagai dos TRF (200, 500 dan 1000 mg/kg), kumpulan kawalan positif
yang dirawat dengan 100 mg/kg butilated hidroksianisol (BHA)
dan kumpulan kawalan yang hanya diberi bahan pelarut (minyak jagung).
Pada hari ke-15, tikus telah dibunuh dan heparnya diasingkan.
RNA jumlah
diekstrak daripada hepar dan seterusnya asai reaksi rerantai polimerase
kuantitatif (qPCR) telah dilakukan untuk menganalisis pengekspresan
gen beberapa isoenzim GST. Protein jumlah hepar juga telah
diekstrak dan pengekspresan protein beberapa isoenzim GST ditentukan
dengan menggunakan kaedah ‘Western blot’. Keputusan kajian menunjukkan
bahawa suplementasi oral TRF menyebabkan peningkatan signifikan
pengekspresan gen dan protein beberapa isoenzim GST yang
bersandarkan dos, berbanding kumpulan kawalan. Kesimpulannya,
suplementasi oral TRF
selama 14 hari pada mencit berupaya menyebabkan
peningkatan pengekspresan gen dan protein beberapa isoenzim GST yang
bersandarkan dos dengan pengekspresan tertinggi diperhatikan pada
mencit yang dirawat dengan 1000 mg/kg TRF. Kata kunci: Glutation S-transferase; hepar; isoenzim; tokotrienol;
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Eur. J. Biochem. 224: 893-899. *Pengarang untuk surat-menyurat; email:
azman.abdullah@ppukm.ukm.edu.my
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