KCa3.1通道参与调控星形胶质细胞糖氧剥夺诱导的内质网应激

摘要

目的:研究KCa3.1在糖氧剥夺诱导的原代星形胶质细胞内质网应激(ERS)中的调控作用.方法:通过构建原代星形胶质细胞糖氧剥夺(OGD)模型,应用cck-8法、免疫荧光技术、western blotting等分子生物学技术研究KCa3.1在OGD引起的原代星形胶质细胞内质网应激中的作用.结果:OGD4h处理后星形胶质细胞内KCa3.1的表达明显上调.OGD处理后星形胶质细胞的细胞活力显著性降低,且具有时间依赖性.给予KCa3.1通道抑制剂TRAM-34可提高OGD 4 h处理后星形胶质细胞的细胞活力,并具有剂量依赖性.OGD处理0.5 h、1h、3h、4h、6h后,原代星形胶质细胞内ERS信号通路被激活,GRP78、p-eIF-2α的表达显著性上调.给予KCa3.1通道抑制剂TRAM-34后,OGD引起的星形胶质细胞内GRP78、p-eIF-2α的上调幅度显著性降低.结论:KCa3.1通道参与了星形胶质细胞内OGD引起的内质网应激通路的激活.%Objective:To evaluate the regulation of KCa3.1 in oxygen and glucose deprivation-induced endoplasmic reticulum stress in primary astrocytes.Methods:The model of oxygen and glucose deprivation in primary astrocytes were constructed to explore the role of Ka3.1 channel on OGD-induced endoplasmic reticulum stress by using cell biology and molecular biology techniques including cck-8,immunofluorescence technique and western blotting.Results:Expression of KCa3.1 protein in astrocytes subjected to OGD 4 h was significantly up-regulated.Cell viability of astrocytes subjected to OGD was significantly time-dependent lower.KCa3.1 channel inhibitor,TRAM-34,could improve the cell viability of astrocytes treated with OGD 4 h,which was dose-dependent.ERS signaling pathway was activated in primary astrocytes at 0.5 h,1 h,3 h,4 h,6 h after OGD.The expression of GRP78 and p-eIF-2α protein was significantly up-regulated.The up-regulation of GRP78 and p-eIF-2α in the TRAM-34 group after OGD treatment was significantly inhibited.Conclusions:KCa3.1 channel is involved in the activation of endoplasmic reticulum stress pathway induced by OGD in primary astrocytes.