Objective To establish a rat model of immunological hepatic injury and to explore the effect of blocking or activating Tim-3 pathway on Th17 cells in the rats. Methods Forty SPF male Wistar rats (body weight 180±20 g) were used in this study. The rat model of immunological hepatic injury was established by injecting 12.5 mg/kg concanavalin A (Con A) through the tail vein once a week for eight weeks. Splenic lymphocytes were isolated. Serum was separated and stored in -20℃. Liver tissue samples were fixed in 4% paraformaldehyde for further study. The splenic lymphocytes were divided into 5 groups: blocking group, blocking control group, activating group, activating control group and ConA control group. The Tim-3 pathway was blocked by anti-Tim-3 while activated by recombinant galectin-9, and cultured 72 h under 37℃, 5% CO₂ environment. The cell culture supernatants were saved at -20℃ for later use. Biochemical analysis was performed to determine the expression of serum ALT, AST and ALB. Morphological changes of the liver were examined by histopathology using HE staining. Immunohistochemical staining was used to detect the expression of IL-17A and ROR-γt proteins in the liver tissues. Enzyme-linked immunosorbent assay (ELISA) was used to assess the IL-17A and IL-6 in the cell supernatants while real time PCR was applied to determine the expression of ROR-γt mRNA in splenic lymphocytes. Results Compared with the control group, inflammatory cell infiltration, hepatic tissue injury and many pseudolobules were easily found in the liver tissue, and immunohistochemical examination showed significantly increased levels of IL-17A and ROR-γt proteins in the liver tissues of the model group. The expressions of IL-17A and IL-6 in the blocking group were increased than that of the blocking control group (P <0.05), while the levels of IL-17A and IL-6 in the activating group were lower when compared with that of the activating control group (P<0.05). Compared with the blocking control group, the real time PCR showed that the expression of ROR-γt mRNA in the blocking group was significantly increased (P<0.05). Conclusions The pathogenesis of immunological hepatic injury is closely related to Th17 cells. Immune-regulating Tim-3 pathways may affect the effect of Th17 cells then further participate in the mechanism of immunological hepatic injury.