Figure 2.
Effects of mTOR activation on HIRI. AT mice were generated by cross-breeding Alb-Cre mice with TSC1floxp/floxp mice. Animals were fed NCD for 16 wk and challenged with HIRI. Rapamycin (1 mg/kg) was administrated intravenously twice, 15 min before ischemia and then immediately before reperfusion. Results are expressed as means ± sem; n = 10 for each group. *P < 0.05 vs. wild-type mice; #P < 0.05 vs. AT mice (Student’s t test). A) Validation of mTOR activation in liver. Liver protein was extracted and used for Western blot. B) Histologic measurement of hepatic lipid content by Oil Red O staining and quantitated by ImageJ software; hepatic triglyceride content and mRNA levels of hepatic lipid metabolism–related genes are expressed as means ± sem. C) TUNEL staining and number of positive hepatocytes shown at low (×10) and high (×40) magnification. Positive staining area and cell numbers were calculated under ×40 magnification. D) Serum levels of ALT and LDH. E) Hepatic levels of inflammatory factors, including MCP-1, TNF-a, and IL-6, were detected by ELISA assay. F) Cleaved caspase 3 was detected by Western blot. β-Actin was used as internal control. Representative results from 3 independent experiments are shown.