黄芪甲苷改善高糖诱导内皮细胞线粒体功能障碍

Astragaloside IV Improves High Glucose-Induced Mitochondrial Dysfunction of Endothelial Cells

目的研究黄芪甲苷对高糖诱导内皮细胞线粒体功能障碍保护作用并探讨其机制。方法体外培养人脐静脉内皮细胞,将细胞分为8组:空白组,高糖组,高糖+阳性药组(维生素C)组,高糖+黄芪甲苷低、高剂量(40、80μmol·L^(-1))组,SRT1720(SIRT1激动剂)组,高糖+SRT1720组,高糖+黄芪甲苷+EX-527(SIRT1抑制剂)组。空白组以5.5 mmol·L^(-1)正常低糖处理24 h,高糖组以30 mmol·L^(-1)高糖处理24 h,SRT1720组用SRT1720处理24 h后正常低糖处理,其余组分别用维生素C,黄芪甲苷,SRT1720和EX-527处理24 h后高糖处理。MitoSOX染色法检测各组细胞线粒体活性氧水平,JC-1染色法检测各组细胞线粒体膜电位水平,免疫荧光检测各组细胞胞核中NF-κB水平,免疫印迹法检测各组细胞Drp1、SIRT1和NF-κB蛋白水平。结果黄芪甲苷可显著降低高糖诱导内皮细胞线粒体活性氧水平,增加线粒体膜电位水平及下调Drp1蛋白表达水平,并能激活SIRT1/NF-κB信号通路。SRT1720同黄芪甲苷的效果一致,而EX-527可取消黄芪甲苷对高糖诱导内皮细胞线粒体功能障碍的改善作用。结论黄芪甲苷可通过激活SIRT1/NF-κB信号通路改善高糖诱导内皮细胞线粒体功能障碍。

Objective To investigate the protective effect of astragaloside IV on mitochondrial dysfunction of endothelial cells induced by high glucose and its mechanism.Methods Human umbilical vein endothelial cells(HUVECs)were cultured in vitro and divided into 8 groups:control group,high glucose group,high glucose+positive drug(vitamin C)group,high glucose+astragaloside IV low and high dose(40μmol·L^(-1)and 80μmol·L^(-1))groups,SRT1720(SIRT1 agonist)group,high glucose+SRT1720 group,high glucose+EX-527(SIRT1 inhibitor)group.The control group was treated with 5.5 mmol·L^(-1)glucose for 24 h,the high glucose group was treated with 30 mmol·L^(-1)high glucose for 24 h,SRT1720 group was treated with SRT1720 for 24 h before treated with 5.5 mmol·L^(-1)glucose.The other groups were respectively treated with vitamin C,astragaloside IV,SRT1720 and EX-527 before treated with high glucose.Mitochondrial reactive oxygen species levels were detected by MitoSOX staining.Mitochondrial membrane potential was detected by JC-1 staining.The level of NF-κB in nucleus was detected by immunofluorescence.Western blotting was used to detect the levels of Drp1,SIRT1 and NF-κB protein.Results Astragaloside IV significantly decreased the levels of mitochondrial reactive oxygen species,increased mitochondrial membrane potential and reduced the level of Drp1 protein expression,and activated SIRT1/NF-κB signaling pathway in endothelial cells induced by high glucose.The effect of SRT1720 treatment was similar to astragaloside IV,while EX-527 treatment abolished the protective effect of astragaloside IV on high glucose-induced mitochondrial dysfunction on HUVECs.Conclusion Astragaloside IV may protect against high glucose-induced mitochondrial dysfunction on endothelial cells via activating SIRT1/NF-κB signaling pathway.