To evaluate the application value of serum enzyme biomarkers of total bile acid (TBA), α-glutathione-S-transferase (α-GST) etc for the early diagnosis of liver injury induced by Rhizoma Menispermi (RM) in rats.Methods：Wistar rats were randomly divided into control group and RM low dose group (22.5 g·kg-1) and high dose (45 g·kg-1) group (n=18). RM 22.5 and 45 g·kg-1 groups were treated with RM decoction by intragastrical administration, while the control group was given distilled water of the same volume once a day for 21 consecutive days. Blood was collected respectively 7, 14, 21 days after RM administration. The levels of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), total bilirubin (TB) and TBA were measured by using automatic biochemical analyzer. The concentration of α-GST, purine nucleoside phosphorlyase (PNP), ornithine carbamyl transferase (OCT) and arginase Ⅰ (Arg Ⅰ) were detected with enzyme-linked immunosorbent assay (ELISA). Liver weight was measured and the pathological changes of liver tissue were observed. Logistic regression and ROC curve were applied to analyze the data of above indexes for evaluating the early diagnostic value of liver injury induced by RM.Results：Compared with that in the control group at the same time point, on day 14 the level of ALP was elevated (P< 0.05) and the level of ALT and liver weight index were increased (P< 0.05) and some spotty and focal necrosis, inflammatory cell infiltration in the liver tissue were demonstrated on day 14 and 21 in RM high dose group. The liver weight index was increased in RM low dose group (P< 0.01). The levels of α-GST, PNP and OCT were elevated on day 7, 14 and 21 (P< 0.05 orP< 0.01), and OCT increased on day 14 and 21, TBA increased only on day 14 in RM high dose group. The areas under the ROC curve (AUC) for serum ALT, AST, α-GST, PNP and Arg Ⅰ were 0.787, 0.483, 0.957, 0.948 and 0.934 respectively. The AUC of the combined examination of ALT and AST was 0.809. The AUC of the combined examination of α-GST, Arg Ⅰ and the AUC of the combined examination of α-GST, Arg Ⅰand PNP were the same of 1.000, higher than the AUC of single index examination.Conclusion：α-GST, PNP and Arg Ⅰ may be used as the biomarkers at the early stage of hepatic injury induced by RM. The combined examination has a higher value for the early liver injury diagnosis.