Figure 1

(A–D) In WT mice, the levels of serum aminotransferases (ALT and AST), TNF-α and IL-6 increased significantly in IR group compared with sham group, and decreased in RPC group compared with IR group (*P < 0.05, vs Sham, ^#P < 0.05, vs IR); In TLR4 KO mice, there were no significant differences between RPC group and IR group. (E,F) Hepatic tissue histological injury extent was detected with H&E staining for light microscopy examination. The area in the photograph signed with black arrows depicted typical pattern of focal necrosis resulted by ischemia. Hepatocyte apoptosis was determined by TUNEL method for immunohistochemical. The area in the photograph signed with black arrows depicted apoptotic positive cells. In WT mice, Areas of necrosis (Magnification: 200×, Scale bar = 200 μm) and percentage of apoptotic cells (Magnification: 400×, Scale bar = 100 μm) were both significantly lower in the RPC group than in the IR group. In TLR4 KO mice, there were no significant difference in liver necrosis and apoptosis between the RPC group and IR group. (G) Western blot analysis of TLR4 protein expression in liver tissues in the wild type and TLR4 KO mice. GAPDH was used as the loading control. (H) RAW264.7 cells were pretreated with 10 ng/ml remifentanil for 1 h and then stimulated by 1 μg/ml LPS for another 6 h, the protein expression of TLR4 was measured by western blot. *P < 0.05 compared to LPS alone. (n = 8 in each group and 3 repeats of each experiment).