Western blot analysis
Total proteins from liver tissue samples were extracted by using the RIPA lysis buffer containing 1% 100 mM PMSF then the concentrations were determined by BCA Protein Assay Kit (Thermo Scientific, 23227, Rockford, IL). Protein levels were detected by Western bolt analysis. Protein samples of 40 μg per lane were separated by a 10% SDS-PAGE gel and transferred onto polyvinylidene fluoride membranes. After blocking with 5% non-fat milk for 1 h, the blots were incubated with different antibodies overnight at 4℃, followed by secondary anti-rabbit (Cell Signaling Technology, #7077, Danvers, MA)or anti-mouse antibodies (Cell Signaling Technology, #7076, Danvers, MA) at room temperature for 1 h on the next day. The ECL Detection Kit (Engreen Biosystem, Beijing, China) was employed to develop the blots. The intensity of the protein bands was assessed by Quantity One software (Bio-Rad, California, USA).