Objective To identify the differential expression of serum proteins in rats after hepatocyte injury induced by ketoconazole (KTZ) and carbon tetrachloride (CCl4). Methods Ninety-six SPF 7-week old male Wistar rats were randomly divided into the KTZ (n = 32), control (n = 32) and CCl4 groups (n = 32). The KTZ group was administered 225 mg/ kg KTZ twice, once a day. The CCl4 group was administered 10 mL/ kg CCl4(V/ V: 30%) once and the control group was given an equal amount of saline once by intragastric administration. At 4, 24, 48 and 72 h after the administration, blood was taken from anesthetized animals and used to prepare serum. The livers were removed and used to prepare pathological sections. Serum proteins from the KTZ and CCl4 groups after hepatocyte injury and from the control group were extracted and differentially expressed proteins were identified by two-dimensional electrophoresis (2D-PAGE) and liquid chromatography-tandem mass spectrometry (LC-MS/ MS). Results The ALT at 24 h after the end of the administration in the KTZ and CCl4 groups was significantly increased with pathological changes in hepatocytes ( P <0. 05). Compared with the control group, four differentially expressed proteins, i. e. apolipoprotein AI, plasma glutathione peroxidase, apolipoprotein E and vitamin D-binding protein were identified in the KTZ and CCl4 groups. Conclusions The four differentially expressed proteins can be used for further studies of potential biomarkers for hepatocyte injury.