PM2.5对人表皮黑素细胞自噬水平的影响

Effects of fine Particulate Matter(PM2.5)on the autophagy in human skin melanocytes

目的研究空气细颗粒物(PM2.5)对人表皮黑素细胞自噬水平的影响。方法收集湖北黄冈市区空气中的PM2.5,分离并提取其水溶性及非水溶性成分,以不同的终浓度处理体外培育人表皮黑素细胞。采用单丹磺酰尸胺(MDC)染色法检测PM2.5对人表皮黑素细胞自噬的影响、采用免疫组化和蛋白印迹法检测PM2.5作用前后人表皮黑素细胞自噬标记性蛋白LC3-II/I的含量表达的差异。结果随着PM2.5水溶性及非水溶性成分浓度的升高,黑素细胞自噬体及自噬标志蛋白LC3-II/I的表达均随之升高,自噬反应明显增强(P<0.01)。蛋白印迹结果表明,在相同浓度下,PM2.5非水溶性成分对LC3-II/I表达的促进作用较PM2.5水溶性成分更强(P<0.01)。结论PM2.5可诱导人表皮黑素细胞发生自噬,并与PM2.5浓度有明显相关性,其机制可能与PM2.5对人表皮黑素细胞的氧化应激损伤有关。

Objective To study the effects of fine particulate matter(PM2.5)on the autophagy in human melanocytes.Methods PM2.5 was collected in Huanggang city.Then the water-soluble extracts and non-water-soluble extracts were isolated and the melanocytes were exposed to W-PM2.5 or NW-PM2.5 with various concentrations.Autophagy of melanocytes was observed by MDC staining,and the expression of LC3-II/I was detected by immunohistochemistry and western blot.Results With the increase of concentration of W-PM2.5 and NW-PM2.5,the expression of melanocyte autophagosome and LC3-II/I increased,and the autophagy response was significantly enhanced(P<0.01).In addition,the results of western blot showed that,at the same concentration,NW-PM2.5 components had a stronger effect on promoting LC3II/I expression than W-PM2.5 components(P<0.01).Conclusion PM2.5 can induce oxidative stress injury and excessive autophagy of melanocytes.