动力相关蛋白1促进高糖诱导的冠状动脉内皮细胞线粒体损伤

Drp1 promotes high glucose⁃induced mitochondrial injury of coronary endothelial cells

目的探讨动力相关蛋白1(Drp1)在高糖诱导的冠状动脉内皮细胞中的表达水平、生物学功能及其对线粒体功能的影响。方法通过实时荧光定量PCR和Western blot实验检测高糖诱导的冠状动脉内皮细胞中Drp1、OPA1及MFN1的表达水平;通过转染Drp1 shRNA表达质粒敲低Drp1的表达,用CCK⁃8实验检测细胞活力,用Annexin V染色实验检测细胞凋亡水平,并通过JC⁃1荧光染色实验和Calcein⁃AM荧光染色实验分别检测线粒体膜电位和线粒体通透性转换孔。结果高浓度葡萄糖处理的冠状动脉内皮细胞中Drp1 mRNA和蛋白的表达显著升高(均P<0.05),细胞活力显著下降(P<0.05),凋亡细胞比例显著升高(P<0.05),线粒体膜电位降低(P<0.05),线粒体通透性转换孔活性升高(P<0.05);敲低Drp1可以增强细胞活力(P<0.05),并显著抑制高糖诱导的细胞凋亡(P<0.05)、线粒体膜电位下降(P<0.05)和线粒体通透性转换孔活性的升高(P<0.05)。结论Drp1在高糖诱导的冠状动脉内皮细胞损伤中表达水平升高,敲低Drp1可以缓解高糖诱导的冠状动脉内皮细胞损伤和线粒体功能损伤。

Objective To explore the expression level,biological function and effect of Drp1 on mitochon⁃drial function in high glucose induced coronary endothelial cells.Methods Real⁃time qPCR and Western blot were used to detect the expression levels of Drp1,OPA1 and MFN1 in coronary endothelial cells induced by high glucose.Drp1 expression was knocked down by transfection of Drp1 shRNA plasmid,cell viability was detected by CCK⁃8 as⁃say,cell apoptosis was detected by Annexin V staining assay,and mitochondrial membrane potential and mitochon⁃drial permeability transition pores were detected by JC⁃1 fluorescence assay and Calcein⁃AM fluorescence assay,re⁃spectively.Results The expression of Drp1 mRNA and protein in coronary endothelial cells treated with high con⁃centration glucose was significantly increased(all P<0.05),the cell viability was significantly decreased(P<0.05),the apoptotic cells was significantly increased(P<0.05),and the mitochondrial membrane potential was decreased(P<0.05).The activity of mitochondrial permeability transition pore was increased(P<0.05).Drp1 knockdown en⁃hanced cell viability(P<0.05),and significantly inhibited high glucose⁃induced cell apoptosis(P<0.05),de⁃creased mitochondrial membrane potential(P<0.05),and increased mitochondrial permeability transition pore activ⁃ity(P<0.05).Conclusion The expression of Drp1 was increased in the coronary endothelial cell injury induced by high glucose.Knocking down Drp1 could alleviate the coronary endothelial cell injury and mitochondrial injury in⁃duced by high glucose.