Hepatoprotective Effects of Punica Granatum Fruit against D-galactosamine Induced Hepatotoxicity in Rats: in Vitro and in Vivo Studies

Objective: Hepatoprotective activity of Punica granatum Linn. (Family: Punicaceae) was evaluated by in vitro and in vivo model. HepG2 cell lines were used for in vitro study and D-Galactosamine (D-GalN) induced hepatic damage model for in vivo evaluation. Methods: Hepatoprotective potential was assessed by measuring serum level of glutamate oxaloacetate transaminase, glutamate pyruvate transaminase, alkaline phosphatase, total bilirubin, and albumin. Enzymatic antioxidant parameters like TBARs (thiobarbituric acid reactive substances), GSH (glutathione), SOD (superoxide dismutase) and CAT (catalase) also evaluated in this study. Results: Rats treated with D-GalN showed a significant increase in serum levels of SGOT (serum glutamate oxaloacetate transaminase), SGPT (serum glutamate pyruvate transaminase), ALP (alkaline phosphatase), bilirubin and TBARs, reflecting liver damage. The in vitro study indicates a beneficial effect of aqueous extract of P. granatum in comparison with methanolic extract, on D-GalN induced toxicity to HepG2 cells. On the basis of in vitro study, aqueous extract was selected for in vivo evaluation. The aqueous extract of P. granatum significantly reduced the elevated serum biomarkers, indicating the recovery of hepatocellular injury. It was also observed that D-GalN induced a significant decrease in GSH, SOD, CAT, protein, and albumin level were increased on the treatment of the rats with aqueous extract of P. granatum. The findings were also confirmed by histopathological studies. Periportal area and extensive hepatocyte damage and haemorrhage are seen in D-GalN treated group. The portal triad with less degree of inflammatory cell infiltration around the bile duct is seen in P. granatum (500 mg/kg)+(D-GalN) treated group. Conclusion: Results of this study revealed that P. granatum fruit could afford a significant protection for the alleviation of hepatic toxicity. Possible mechanism may involve its action against oxidative stress.