IN VITRO AND IN VIVO PROTECTIVE EFFECTS OF AMBREX, A POLYHERBAL FORMULATION, AGAINST METHOTREXATE INDUCED DAMAGES IN HEPATIC CELLS
Objective: To evaluate the hepatoprotective effect of Ambrex, a poly herbal formulation against methotrexate (MTX) induced hepatotoxicity in Swiss albino mice as well as in Chang liver cell lines.
Methods: Ambrex was exposed to MTX intoxicated chang liver cells and cells were harvested for studying the gene expressions of Dihydrofolate reductase (DHFR), B-cell lymphoma 2 (BCL2) and Bcl-2-associated X protein (BAX). In in vivo study, Ambrex was administered orally for a period of 7 days at two dose levels (250 and 500 mg/kg b. wt) and MTX (20 mg/kg b. wt, i. p) was injected one hour after the last test drug administration. Protective effect of Ambrex was evaluated by measuring aspartate transaminase (SGOT), alanine transaminase (SGPT), alkaline phosphatase (ALP), Î³â€“glutamyl transferase (Î³GT) and total bilirubin. Its effect on superoxide dismutase (SOD), catalase (CAT), reduced glutathione (GSH) and lipid peroxide (LPO) was also determined.
Results: Data revealed that Ambrex was able to restore the levels of antioxidants such as SOD, Catalase, and Glutathione to near normal and reduced the elevated plasma levels of SGOT, SGPT, ALP, Î³â€“GT and total bilirubin. It also inhibited the formation of hepatic malondialdehyde induced by MTX. In vitro studies revealed that Ambrex protected MTX induced hepatotoxicity at the dose of 50 and 500ng/ml. Further, mRNA expression also illustrated that Ambrex inhibited the over expression of BAX and suppressed BCL2 and DHRF expressions.
Conclusion: Results suggest that Ambrex has potent hepatoprotective effect which was evident from both in vivo and in vitro results.
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