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

 

Received: 29 March 2018/Accepted: 30 July 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; TRF

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*Corresponding author; email: azman.abdullah@ppukm.ukm.edu.my

 

 

 

 

 

 

 

 

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