Fig. 4

Berberine reverses neuroinflammation in the prefrontal cortex and hippocampus by inhibiting the NLRP3 inflammasome. a Representation of the timing of BrdU injection and illustration of an image of the target of PFC and hippocampus. b Immunofluorescence of NLRP3 inflammasome in the dentate gyrus subregion of the hippocampus. Scar bar, 100 µm. c Immunofluorescence of NLRP3 inflammasome in the CA1 subregion of the hippocampus. Scar bar, 100 µm. d Immunofluorescence of NLRP3 inflammasome in the PFC. Scar bar, 100 µm. e NLRP3 + cells (green) and Iba1 + cells (red) in DG of hippocampus, CORT increased NLRP3 + /Iba1 + cells in DG (One-way ANOVA, F (3, 8) = 18.63, P = 0.0006; Dunnett’s multiple comparisons test, CORT vs. Control, P = 0.0002, CORT vs. BBR100, P = 0.0112, CORT vs. BBR200, P = 0.0047, n = 3 in each group). f NLRP3 + cells (green) and Iba1 + cells (red) in CA1 of hippocampus, CORT increased NLRP3 + /Iba1 + cells in CA1 (One-way ANOVA, F (3, 8) = 12.28, P = 0.0023; Dunnett’s multiple comparisons test, CORT vs. Control, P = 0.0021, CORT vs. BBR100, P = 0.7604, CORT vs. BBR200, P = 0.0115, n = 3 in each group). g NLRP3 + cells (green) and Iba1 + cells (red) in PFC, CORT increased the NLRP3 + /Iba1 + cells in PFC (One-way ANOVA, F (3, 8) = 141.0, P < 0.0001; Dunnett’s multiple comparisons test, all comparisons P < 0.0001, n = 3 in each group). h The western blotting of NLRP3/Caspase-1/ASC in the PFC, the CORT mice showed a significant difference in the NLRP3 signaling pathway (Two-way ANOVA, row factor F (2, 36) = 10.30, P = 0.0003, column factor F (3, 36) = 24.85, P < 0.0001; Dunnett’s multiple comparisons test, all comparisons P < 0.0001, n = 3 in each group). i The western blotting of NLRP3/Caspase-1/ASC in the hippocampus, the CORT mice showed a significant difference in the NLRP3 signaling pathway (Two-way ANOVA, row factor F (2, 41) = 25.79, P < 0.0001, column factor F (3, 41) = 37.04, P < 0.0001; Dunnett’s multiple comparisons tests, all comparisons P < 0.0001, n = 3 in each group). Bar graphs show the mean ± SEM; *P < 0.05, **P < 0.01, ***P < 0.001. CON control, CORT corticosterone, BBR berberine, ANOVA analysis of variance, NLRP3 NOD-like receptor thermal protein domain associated protein 3, qPCR quantitative reverse transcription polymerase chain reaction, PFC prefrontal cortex