高糖诱导的大鼠雪旺细胞凋亡的流式细胞术

探讨流式细胞术在检测高糖诱导的早期雪旺细胞凋亡中应用。选用培养大鼠的雪旺细胞,将细胞分为正常对照组(5.6mmol/L葡萄糖),高糖组(50mmol/L葡萄糖),高渗出组(50mmg/L甘露醇),培养7天后,收集细胞。48h后应用流式细胞术采用Annexin-V/PI检测法和JC-1检测早期细胞的细胞膜凋亡率和线粒体膜电位变化。结果表明:高糖与正常糖浓度处理的雪旺细胞比较,高浓度葡萄糖诱导雪旺凋亡率明显高于正常葡萄组的凋亡率,线粒体膜电位明显高于正常对照组。高渗透组与正常对照组凋亡率及线粒体无明显升高。通过采用流式细胞细胞术结合单克隆抗体,能够快速、灵敏和准确检测雪旺细胞在细胞膜的通透性改变和线粒体膜电位变化。为研究雪旺细胞凋亡提供客观、直接、准确的方法。

Application of flow cytometry to detect high-glucose induced rats' Schwann cells' apoptosis

To investigate the effect of flow cytometry in detecting high-glucose induced Schwann cells early apoptosis.Using cultured ratsi Schwann cells and dividing them into control group (5.6mmol/L glucose),high glucose group (50mmol/L),and high leakage group (50mmg/L Mannitol).Collecting cells after 7 days.After 48 hours using flow cytometry by Annexin-V/PI and JC-1 to detect early cellsi membrane apoptosis rate and mitochondrial membrane potential change.The results showed that the apoptosis rate and mitochondrial membrane potential of high glucose group were higher than those of control group.However,in high leakage group there were no significant differences compared with the control group.Through flow cytometry binding monoclonal antibody,change of membranous permeability and mitochondrial membrane potentials of Schwann cells could be tested fast,sensitive and accurately.This will provide objective,direct and accurate methods to investigate Schwann cells' apoptosis.