Fig. 2

AICAR increases the engulfment of HMGB1 and reduces inflammation in mouse BMDMs and human PBMC-derived macrophages. A rHMGB1 (his tag) content in BMDMs treated with L-OHP (1 μM). Cells were co-cultured with rHMGB1 (his tag) (10 nM) for 1 h after L-OHP administration (n = 3). B Phosphorylation of AMPK in BMDMs treated with L-OHP (1 μM) (n = 3). C Phosphorylation of AMPK in mice DRG. DRG samples were collected on the 14th day after the first administration of L-OHP (3 mg/kg, i.p.) or vehicle (n = 3). D The general scheme of experiments in vitro. Confocal microscopy images (red) E or content F of rHMGB1 (his tag) in BMDMs (n = 3). Scale bar: 20 μm. Phosphorylation of AMPK in BMDMs G or PBMC-derived macrophages I treated with AICAR (300 μM) for 15 min (n = 3). rHMGB1 (his tag) content in BMDMs H or PBMC-derived macrophages J (n = 3). (K) mRNA level of TNF-α, IL-6 and IL-1β in BMDMs (n = 3). Cells were pretreated with AICAR (300 μM) or AMPK inhibitor Compound C (CC, 20 μM) for 15 min and then co-cultured with rHMGB1 (his tag) (10 nM) for 1 h (E, F, H, I and J). A significant difference was revealed following unpaired Student’s t-test (C, G and I) or one-way ANOVA (A, B, F, H, J and K) (*p < 0.05, **p < 0.01 and ***p < 0.001 vs. Control; ^#p < 0.05, ^##p < 0.01 and ^###p < 0.001 vs. AICAR group; Bonferroni post hoc tests)