Inflammatory activation of BV2 cells induced by manganese involving with mitophagy
Objective:To investigate whether manganese-induced inflammatory activation in BV2 cells is associ-ated with mitophagy.Methods:Mouse microglia cells BV2 were exposed to different concentrations of manga-nese(0 μmol/L,50 μmol/L and 100 μmol/L)for 12 h,and 1 μg/mL lipopolysaccharide(LPS)was used as a posi-tive control group.Cell vialibility was detected by Alarmarblue assay and changes of lysosomal number and fluo-rescence intensity were detected by fluorescent probe;autophagic changes were observed by transmission elec-tron microscopy;the extent of lysosomal and mitochondrial fluorescence co-localization was observed by confo-cal microscopy;western blotting was performed to detect the expression levels of cellular inflammatory proteins NLRP3,autophagy-related proteins(p62,LC3-Ⅱ/Ⅰ),and mitochondrial outer membrane protein VDAC1 after 12 h of exposure to different concentrations of manganese.Mitophagy inhibitor Bafilomycin A1 pretreatment veri-fied the effect of manganese exposure on autophagic flow and the aforementioned inflammatory hallmarks pro-tein expression.Results:When BV2 cells were exposed to manganese at concentrations greater than 50 μmol/L,the BV2 cell survival rate was decreased,NLRP3 expression was increased(P<0.01),and the number of lyso-somes and their co-localization with mitochondria significantly were increased(P<0.05).The protein expression levels of VDAC1 and autophagy marker protein LC3-Ⅱ/Ⅰ were decreased and p62 was increased in the manga-nese-exposed group(P<0.05).Bafilomycin A1 pretreatment significantly reversed the autophagy marker protein expression except p62(P<0.05).Conclusion:Manganese induces inflammatory activation in BV2 cells and in-creases mitophagy at 50-100 μmol/L exposure dose.Inhibition of mitophagy by Bafilomycin A1 can promote manganese exposure-induced inflammatory activation in BV2 cells.
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维普
