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

KCa3.1 Channel was involved in the Regulation of Oxygen and Glucose Deprivation-induced Endoplasmic Reticulum Stress in Astrocytes

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:OGD 4 h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、1 h、3 h、4 h、6 h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, imrnunofluorescence technique and western blotting. Results： Expression of KCa3. I 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-elF-2α protein was significantly up-regulated. The up-regulation of GRP78 and p-elF-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.