Regulation of Acrylamide Induced µ-Class Glutathione-S-Transferases in Mice Testis by the Treatment of Hybanthus Enneaspermus Active Principles.

ABSTRACT

The present study was aimed to analyse the effect of acrylamide and Hybanthus enneaspermus leaf extract active principles on mice testis glutathione-s-transferases (GST; EC 2.5.1.18). These enzymes play a role in biotransformation of electrophilic compounds that cause damage to cells by conjugating with the substrate glutathione. Hybanthus enneaspermus, a spade flower, is an erect shrub of violaceae family, having free radical scavenging activity. Acrylamide is a known neurotoxicant that cause damage to almost all cells including liver, testis, brain and kidney. The GSTs purified from mice testis using glutathionyl linked agarose affinity chromatography were analyzed by using SDS-PAGE and were resolved into four sub units i.e. Yc, Yb, Yβ &Yδ. Also these subunits expression were confirmed by western blot analysis. During experimentation to analyze the effect of Hybanthus enneaspermus active principle (HE) mice were subjected to both acrylamide (AC) and also mixture of HE and AC. This exposure significantly altered the specific activity of mice GSTs in testis. Polyclonal antibodies produced against purified GSTs of mice testis on immunoblot analysis showed significant increase of μclass GSTs (Yb & Yβ) based on dose and time dependent manner. Therefore the present research of Hybanthus enneaspermus treatment on mice testis showed that, regulation of synthesis of μ-GSTs was depending on the dose of acrylamide concentration and also the active principles of HE. Hence it is proposed that μ-GSTs may be used as tumour markers for testis carcinoma, since their production is variable due to the increased dose concentration of synthetic chemical acrylamide and its regulation by plant product, HE.