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目的观察体积调节性氯离子通道(volume-regulated anion channel,VRAC)阻滞剂NPPB(5-nitro-2-(3-phenyl progy(amino)-benzoic acid)对低渗条件下小胶质细胞活化及其炎性分泌的作用。方法小胶质细胞系BV2细胞传代培养,分为对照组、低渗组、低渗加NPPB组,在低渗干预1h后给予正常培养基继续培养,在3、6、18和24h收取细胞和细胞培养基;用ELISA法测定各组培养基中IL-6和TNF-α的浓度变化。结果与对照组比较,低渗干预后BV2细胞分泌的IL-6和TNF-α增加,6h时BV2细胞分泌的IL-6和TNF-α达到最高峰(p<0.01);低渗加NPPB可显著降低IL-6和TNF-α的分泌(p<0.01),在6h时抑制效果最为显著。结论低渗条件可诱导BV2细胞活化及炎性分泌增加,VRAC阻滞剂NPPB可减少低渗条件下BV2细胞的活化和分泌,提示VRAC在低渗诱导的小胶质细胞活化及炎性因子分泌中发挥了重要作用。
Objective To observe the effects of 5-nitro-2- (3-phenyl progy (amino) -benzoic acid), a volume-regulated anion channel (VRAC) inhibitor, on microglial activation And its role in inflammatory secretion.Methods BV2 cells were subcultured and divided into control group, hypotonic group and hypopotentiation plus NPPB group.After 1 h of hypotonic intervention, normal culture medium was given to continue culture, 6, 18, and 24 h after culture, and the concentrations of IL-6 and TNF-α in each group were measured by ELISA method.Results Compared with the control group, the secretion of IL-6 and BV- The levels of IL-6 and TNF-α secreted by BV2 cells peaked at 6h (p <0.01) after hypoxia and NPPB treatment (p <0.01) 6h.Conclusion The hypotonic conditions can induce BV2 cell activation and inflammatory secretion increased, VRAC blocker NPPB can reduce BV2 cell activation and secretion under hypotonic conditions, suggesting VRAC in the hypotonic induced microglia Cell activation and secretion of inflammatory cytokines play an important role.