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Protective Effect of Trigonella foenum-graecum and Foeniculum vulgare Mature Leaf Against t-BHP Induced Toxicity in Primary Rat Hepatocytes

Abstract

Pankaj Singh,Poonam Kakkar,Ram Lakhan Singh*

Objective: Trigonella foenum-graecum (TFG), is used to treat diabetes, liver and spleen diseases, whereas Foeniculum vulgare (FV), is used to cure liver disorder, cardiovascular diseases, cancer, aging etc. in Ayurvedic system. Both plants have variety of pharmacological activities, including anticancer, antidiabetic, antihepatotoxic, antibacterial activities etc. The present study was carried out to explore and compare the hepatoprotective activities of ethanolic extracts of mature TFG and FV leaf against tert-butyl hydroperoxide (t-BHP) induced cell death in primary rat hepatocytes. Methods: Hepatocytes were isolated from liver of overnight fasted rat after subjecting it to two stage collagenase perfusion with HEPES {4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid} buffer. Only preparations with cell viability greater than 95% were used for subsequent experiments. Cell viability was determined by MTT {3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide} assay, whereas hepatoprotective activities of plant extracts were determined by estimation of superoxide dismutase and catalase activities, lipid peroxidation, total glutathione content and nitric oxide release against t-BHP induced oxidative stress. Results: Treatment of TFG and FV leaf extracts (1.0 to 7.5 μg) significantly reduced the impact of t-BHP (250 μM) induced toxicity. TFG and FV leaf extracts showed 2.3 and 2.7 fold (P<0.001) increase in superoxide dismutase (SOD) activity, 3.4 fold and 2.2 fold (P<0.001) increase in catalase (CAT) activity at concentration of 5 μg whereas maximum glutathione (GSH) content was restored at concentration of 2.5 μg and 5.0 μg, respectively as compared to stressed cells. Both the leaf extracts were found to decrease the lipid peroxidation (P<0.001) and inhibit NO release (P<0.001) at concentration of 5.0 μg in cells as compared to stressed cells. Conclusion: The results suggest cytoprotective, antioxidant and hepatoprotective properties of TFG and FV leaf extracts which may be used in the form of dietary component and also in formulations against liver diseases.

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