Figure 5.

Metformin inhibited HMGB1-induced p38 phosphorylation in mouse peritoneal macrophages. Mouse peritoneal macrophages from Balb/c mice were incubated at 37 °C for 1 h with 1 μg/ml full-length HMGB1 or 1 μg/ml HMGB1-ΔAT in the absence or presence of 3 mm metformin. HMGB1 or HMGB1-ΔAT was preincubated without or with metformin at 4 °C for 24 h before the addition to cells. Cell lysates were then analyzed by immunoblotting for phosphorylated and total p38 and for phosphorylated and total AMPK. A, typical photos of immunoblots. Data shown are representative of four independent experiments with similar results. B and C, ratios of phosphorylated/total p38 (B) and phosphorylated/total AMPK (C) were calculated from densitometry analysis. B and C, results were analyzed as the percentage of control group (HMGB1 alone or HMGB1-ΔAT alone). Data are presented as mean ± S.E. (n = 4). *, p < 0.05 versus control group.