缺氧诱导因子1α对小胶质细胞M1极化的影响及其机制研究

Effect of hypoxia-inducible factor 1α on microglia M1 polarization and its mechanism

目的 研究缺氧诱导因子1α(HIF-1α)对小胶质细胞M1极化的影响及其影响机制。方法 将小胶质细胞(BV-2细胞)随机分为6个组:Control组和10、50、100、200、500μg/L重组HIF-1α蛋白刺激处理组。采用荧光共聚焦法观察各组BV-2细胞的形态变化;采用免疫蛋白印迹法定量分析重组HIF-1α蛋白刺激处理后NF-κB p65、p-STAT1和TRAF6蛋白含量变化。结果 与对照组相比,重组HIF-1α蛋白刺激后小胶质细胞胞体变大,呈圆形或吞噬状,突起变粗变短;胞内NF-κB p65、TRAF6蛋白较对照组显著增加,不同浓度重组HIF-1α蛋白刺激处理组对比结果有显著差异。结论 HIF-1α可刺激小胶质细胞M1极化,不同浓度重组HIF-1α蛋白刺激处理有量效关系,其机制可能与通过TLR4/Myd88/NF-κB通路调节TRAF6和NF-κB活化有关。

Objective To study the effect of hypoxia-inducible factor 1α(HIF-1α) on microglia M1 polarization and its mechanism.Methods Microglia(BV-2 cells) were randomly divided into six groups:control,10,50,100,200 and 500 μg/L recombinant HIF-1α groups.Morphological changes of BV-2 cells were observed by fluorescence confocal microscopy.Changes of nuclear factor(NF)-κB p65,p-STAT1 and TRAF6 protein contents after recombinant HIF-1α protein stimulation were quantitatively analyzed by Western blot.Results Compared with the control group,microglia cells stimulated by recombinant HIF-1α protein became larger,round or phagocytic,and their processes became thicker and shorter.Intracellular NF-κB p65 and TRAF6 proteins were significantly increased compared with the control group,and the result of groups with varying concentrations of recombinant HIF-1α protein stimulation were significantly different.Conclusions HIF-1α can stimulate microglial M1 polarization and there was a dose-effect relationship between different concentrations of recombinant HIF-1α,which may be related to the regulation of TRAF6 and NF-κB activation through the TLR4/Myd88/NF-κB pathway.