黄芪甲苷上调HIF-1α表达促进氧糖剥夺/再灌注胚鼠海马神经干细胞增殖和分化的作用

Astragaloside up-regulates HIF-1αexpression to promote the proliferation and differentiation of NSCs in the hippocampus of OGD/R embryonic mice

目的:探讨黄芪甲苷对离体氧糖剥夺/再灌注(OGD/R)胎鼠海马神经干细胞(NSCs)自我更新、增殖和分化的作用及其机制。方法:分离、培养、鉴定胎鼠海马NSCs。建立NSCs体外OGD/R模型,通过Western blot和免疫荧光双标方法验证黄芪甲苷对NSCs的增殖与分化的作用及可能的机制。结果:筛选出黄芪甲苷促进NSCs增殖分化的最佳浓度为0.25μg/mL,作用的最佳时间点为24h。黄芪甲苷可促进海马NSCs中BrdU的表达,BrdU、Nestin、Tuj-1、Vimentin阳性细胞数的表达增多。与正常组比较,氧糖剥夺显著增加了NSCs中HIF-1α蛋白含量(P<0.01);与模型组比较,黄芪甲苷组HIF-1α蛋白含量显著增多(P<0.01)。结论:黄芪甲苷上调HIF-1α信号通路,增强氧糖剥夺NSCs增殖与分化的能力,而促进脑缺血损伤的结构和功能的修复。

Objective:To investigate the effect and mechanism of astragaloside on the self-renewal,proliferation and differentiation of hippocampal neural stem cells(NSCs)in vitro after oxygen-glucose deprivation/reperfusion(OGD/R)rats.Methods:NSCs in hippocampus of fetal rats were isolated and identified.Establish the in vitro OGD/R of NSCs,and to verify the effect and possible mechanism of astragaloside on proliferation and differentiation of NSCs by Western blot and immunofluorescence double-label method.Results:The optimal concentration of astragaloside for promoting proliferation and differentiation of NSCs was 0.25μg/mL,and the optimal time point of action was 24h.Astragaloside could promote the expression of BrdU in hippocampal NSCs,the number of BrdU,Nestin,Tuj-1,Vimentin positive cells were increased.Compared with the normal group,the protein expression of HIF-1αwas significantly enhanced by OGD(P<0.01).Compared with the model group,the expression of HIF-1αwas significantly promoted in astragaloside group(P<0.01).Conclusion:Astragaloside up-regulates HIF-1αsignaling pathway,enhances the ability of proliferation and differentiation of NSCs deprived by oxygen glucose,and promotes the repair of structure and function of cerebral ischemia injury.