Abstract: This work aimed to investigate protective effect of caffeic acid on CCl₄-induced BRL hepatocyte injury. The experiment was divided into control group, CCl₄ model group (100 mmol/L CCl₄ injury for 3 hours) and CA pretreated group (pretreated with 0.2, 0.4 and 0.8 mg/mL CA in DMEM for 4 hours, followed by CCl₄ exposure for 3 hours). The activities of aspartate aminotransferase (AST), alanine aminotransferase (ALT) and lactate dehydrogenase (LDH) were detected by automatic biochemical analyzer. The levels of reactive oxygen species (ROS), cytochrome C (Cyt c) and 8-hydroxydeoxyguanosine (8-OHdG) were measured by the kit. The expression levels of nuclear factor E2-related factor 2 (Nrf2), glutathione reductase (GSR), quinine oxidoreductase (NQO1) and superoxide dismutase (SOD) were detected by qRT-PCR. Significant inhibitory effects of CA were observed on the CCl₄-induced elevation of AST, ALT, LDH, ROS, Cyt c and 8-OHdG in culture medium (P<0.05). Compared with the control group, no significant differences of the mRNA expression levels of Nrf2, GSR, NQO1 and SOD gene were found in the model group (P>0.05). However, the mRNA expression levels of all detected genes were significantly increased in CA-treated group compared with the control group and a dose-dependent manner was observed. In conclusion, CCl₄ exposure induced oxidative stress in BRL hepatocytes and CA inhibited the CCl₄-induced BRL hepatocyte damage by activating the Nrf2/ARE signaling pathway and up-regulating the mRNA levels of antioxidant genes.