Glycyrrhizin attenuates HMGB1-induced hepatocyte apoptosis by inhibiting the p38-dependent mitochondrial pathway

Figure 1 High-mobility group box 1 enhances hepatocyte apoptosis via a mitochondrial pathway. A: Huh-BAT cells were treated with high-mobility group box 1 (HMGB1) (0, 0.001, 0.01, 0.1, 0.5, 1, 5 and 10 μg/mL) for 6 h. Apoptosis was quantified using an APO Percentage apoptosis assay kit. Data are expressed as the mean ± SD of three individual experiments. ^aP < 0.05, vs HMGB1 0 μg/mL; B: Huh-BAT cells were treated with 10 μg/mL of HMGB1 for the indicated time periods. ^cP < 0.05, vs 0 h; C: Huh-BAT cells were treated with HMGB1 (0 μg/mL, 0.1 μg/mL, 0.5 μg/mL, 1, 5 μg/mL and 10 μg/mL) for 6 h (left column), or with 10 μg/mL of HMGB1 for the indicated time periods (right column). Cells were lysed at the indicated time points, and immunoblot analysis was performed using anti-caspase 8 and anti-caspase 3 antibodies. Mitochondrial and cytosolic extracts were also isolated, and equivalent amounts of cytosolic protein were immunoblotted with an anti-cytochrome c antibody. Immunoblot analysis using an anti-actin antibody was performed as a control for protein loading.