Fig. 1

RFAs inhibited NLRP3 inflammasome. Mouse BMDMs were pretreated by diacerein (40 μM) and rhein (40 μM) for 30 min and then stimulated with LPS (100 ng/ml) for 4 h and nigericin (2.5 μM) for 2 h. IL-1β (A) and TNF-α (B) in cell culture supernatant were detected by ELISA (n = 3/group). Mouse BMDMs were pretreated by RFAs (40 μM rhein/40 μM diacerein, 40 μM emodin, 40 μM aloe emodin, 40 μM 1,8-dihydroxyanthraquinone, 20 μM chrysophanol, 4 μM emodin methyl ether, 4 μM anthraquinone) for 30 min and then stimulated with LPS (100 ng/ml) for 4 h and nigericin (2.5 μM) for 2 h. IL-1β (C) and TNF-α (D) in cell culture supernatant were detected by ELISA (n = 3/group), the cleavage of procaspase-1 (E) was detected by western blot, the ASC specks (F, G) were detected by immunofluorescence. Mouse BMDMs were pretreated with some important pharmaceutical ingredients derived from artemisia capillaris (green columns), gardenia (blue columns) and rhubarb (red columns) for 30 min and then stimulated with LPS (100 ng/ml) for 4 h. Nigericin (2.5 μM) was added for 2 h. IL-1β (H) and TNF-α (I) in cell culture supernatant were detected by ELISA (n = 3/group). Mouse BMDMs were pretreated by YCHT (5 μl/ml), artemisia capillaris (5 μl/ml), gardenia (5 μl/ml) and rhubarb (5 μl/ml) for 30 min and then stimulated with LPS (100 ng/ml) for 4 h and nigericin (2.5 μM) for 2 h. IL-1β (J) and TNF-α (K) in cell culture supernatant were detected by ELISA (n = 3/group). Data are presented as mean ± SEM. For multiple comparisons, one-way ANOVA coupled with LSD’s post hoc testing was performed. &: p < 0.05; &&: p < 0.01; &&&: p < 0.001. ANOVA, analysis of variance; ASC, apoptosis-associated speck-like protein containing CARD; BMDMs, bone marrow-derived macrophages; DAPI, 4′,6-diamidino-2-phenylindole; ELISA, enzyme linked immunosorbent assay; IL-1β, interleukin-1 beta; LPS, lipopolysaccharide; TNF-α, tumor necrosis factor-alpha