The study aimed to explain the hepatoprotective action of Cyperus rotundus methanolic extract in vitro and in vivo (albino male mice). An interaction between the extract and Glucophage drug were also made to evaluate the in vivo modulating effects of the extract. The hepatoprotective evaluations included assessment of malondialdyde (MDA) and catalase (CAT) activities and liver function enzymes (aspartate aminotransferase; AST and alanine aminotransferase; ALT and alkaline phosphatase; ALP) in blood serum, as well as histopathological estimation of liver tissue, in mice after administration of the drug Glucophage and cyperus rotundus methanolic extract alone or with interaction. The results indicated that the plant extract (200mg/kg) decreased MDA activity and increase significantly activity of catalase in comparison to positive and negative controls. Also Cyperus rotundus methanolic increase the level of ALT, AST and ALP. The drug Glucophage was contributed to increased MDA activity and decreased the activity of catalase in comparison to positive and negative controls in addition to increase the activity of ALT, AST and ALP. The interaction between the extract and glucophage increased both MDA and catalase activity in comparison to control group. In addition to increase the liver function enzymes ALT, AST and ALP. The histopathological activity represented that histopathological section in the liver of mouse treated with Cyperus rotundus extraction showed scattered degenerated cells while histopathological section in the liver of mouse treated with Glucophage showed normal architecture with few pyknotic nuclei. Histopathological section in the liver of mouse treated with Cyperus rotundus and Glucophage showed multiple aggregation mononuclear cells around central vein.