Fig. 6

miR-146a-5p inhibits neuronal NLRP3 inflammasome in neurons after IH by targeting HIF1α and regulating mtROS expression. (a) Western blot for HIF1α was performed on mice exposed to IH for 8weeks. (b) Protein expression was normalized to β-actin and expressed as a fold change in control. Data are expressed as mean ± SD (n = 4) and analyzed using student’s t test. *P < 0.05, **P < 0.01, and ***P < 0.001. (c) The expression analysis of HIF1α mRNA was significantly increased after transfection of plasmids with high expression of HIF1α. Data are expressed as mean ± SD and analyzed using student’s t test. (d-e) The mRNA expression levels of HIF1α, NLRP3, IL-1β, IL-18 was detected in cultured neurons or culture medium of the three groups (neurons treated with IH for 12 h (IH group), neurons treated with IH for 12 h and transfected of miR-146a-5p mimics (IH + miR-146a-5p mimics group), and neurons treated with IH for 12 h, and transfected of miR-146a-5p mimics and plasmids with high expression of HIF1α (IH + miR-146a-5p mimics + HIF1α group)). Data are presented as mean ± SD and analyzed using one-way ANOVA with Tukey’s post hoc test. (f) The level of mitoSOX in the neurons in the three groups was observed by IF staining, which reflected the level of mtROS. Scale bar: 50 μm. Data are presented as mean ± SD and analyzed using one-way ANOVA with Tukey’s post hoc test. *P < 0.05, **P < 0.01, and ***P < 0.001