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Background:Rhizoma Dioscoreae bulbiferae could cause liver damage,which limited its application in the clinic.Aims and Objectives:To explore the mechanism of hepatotoxicity of Rhizoma Dioscoreae bulbiferae in mice.Materials and Methods:In the present study,the water extraction of Rhizoma Dioscoreae bulbiferae(W.E.R)was administrated via intragastrical with Low(19.6 g/kg),Middle(28.0 g/kg),and High(40.0 g/kg)dose in mice.At each time point 14 days,21 days,and 28 days,the body weight,liver coefficient,indexes of liver function alkaline phosphatase(ALP),alanine aminotransferase(ALT)and aspartate transaminase(AST),various biochemical biomarkers of liver tissue and mitochondria were detected and analyzed.Results:We found that W.E.R could decrease the body weight,increase the liver coefficient and the expression levels of indexes of liver function in mice.Next,we investigated that W.E.R could increase the expression levels of reactive oxygen species(ROS)and malondialdehyde(MDA),decrease the expression levels of adenosine triphosphate(ATP)and improve the enzyme activities of total superoxide dismutase(SOD).Third,we found that W.E.R could increase the enzyme activities of manganese-superoxide dismutase,decrease the enzyme activities of sodium-potassium adenosine triphosphatase(Na+-K+-ATPase)and calcium-magnesium adenosine triphosphatase(Ca 2+-Mg 2+-ATPase).Finally,we analyzed that there were significant negative correlations between body weight,expression level of ATP,activity of Na+-K+-ATPase,activity of Ca 2+-Mg 2+-ATPase and time,dose.There were significant positive correlations between liver coefficient,ALP,ALT,AST,expression levels of ROS,MDA and time,dose.Conclusion:All the results above indicated that W.E.R could cause the hepatotoxicity based on the oxidative damage in mice,which and mitochondria might be the main targets.