黄芪甲苷抗高糖受损内皮祖细胞氧化应激损伤的机制研究  

Study on the Anti-inflammation Effect of Astragalus Membranaceus Fructus on High Glucose-Induced Endothelial Cell Oxidative Stress Damage in Human

作　　者：陈艳[1] 高入春 广可颜 孙忞 彭光霖 瞿文静 贺有缘 赵思佳 邹晓玲[1] 熊武 Damage in Human CHEN Yan;GAO Ruchun;GUANG Keyan;SUN Min;PENG Guanglin;QU Wenjing;HE Youyuan;ZHAO Sijia;ZOU Xiaoling;XIONG Wu(First Affiliated Hospital of Hunan University of Chinese Medicine,Changsha Hunan 410007,China;School of Integrated Chinese and Western Medicine,Hunan University of Chinese Medicine,Changsha Hunan 410208,China)

机构地区：[1]湖南中医药大学第一附属医院,湖南长沙410007 [2]湖南中医药大学中西医结合学院,湖南长沙410208

出　　处：《中医药导报》2025年第1期8-12,19,共6页Guiding Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金面上项目(82374276);湖南省科技厅临床医疗技术创新引导项目(2021SK51412);湖南省卫生健康委科研计划项目(202204034510);湖南省自然科学基金科卫联合项目(2021JJ70033)。

摘　　要：目的:研究黄芪甲苷(AS-Ⅳ)对高糖诱导的内皮祖细胞(EPCs)氧化应激损伤的保护机制,为AS-Ⅳ的临床开发应用奠定基础。方法:取足月新生儿脐带血,采用密度梯度离心法分离出单个核细胞,采用Dil-ac-LDL和FITC-UEA-1双荧光染色法鉴定出EPCs,将鉴定成功的人EPCs分别用30 mmol/L的葡萄糖干预120 h,制成高糖诱导氧化应激损伤细胞模型,实验分为黄芪甲苷+高糖受损EPCs组、黄芪甲苷+高糖受损EPCs+ML385(Nrf2抑制剂)组、高糖受损EPCs组和正常EPCs组(5 mmol/L的葡萄糖干预120 h),用流式细胞术检测活性氧(ROS)水平,蛋白免疫印迹法(Western blotting)检测氧化应激相关蛋白的表达和Nrf2、Keap1及NFE2L2/ARE依赖性信号通路蛋白的表达。结果:与正常EPCs组比较,高糖受损EPCs组细胞内ROS水平显著升高,GSH-Px、CAT、SOD活力均明显降低,LDH活力明显升高,Nox2、Keap1表达显著升高,Nrf2、HMOX1、xCT、GPX4表达降低,差异均有统计学意义(P<0.05);与高糖受损EPCS组比较,黄芪甲苷+高糖受损EPCs组细胞内ROS水平降低,GSH-Px、CAT、SOD活力升高,LDH活力降低,Nox2、Keap1表达降低,Nrf2、HMOX1、xCT、GPX4表达升高,差异均有统计学意义(P<0.05)。黄芪甲苷+高糖受损EPCs+ML385组各项检测指标与高糖受损EPCS组比较,差异无统计学意义(P>0.05)。结论:高糖能诱导EPCs出现氧化应激损伤,而黄芪甲苷能通过介导NFE2L2(Nrf2)/ARE信号通路发挥抗氧化应激作用,保护高糖诱导氧化应激损伤的内皮祖细胞。Objective:To study the protective mechanism of astragaloside IV(AS-IV)against oxidative stress damage of endothelial progenitor cells(EPCs)induced by high glucose,and to lay a foundation for further research on clinical development and application of AS-IV.Methods:Term newborn umbilical cord blood was collected.EPCs were separated by density gradient centrifugation method,and identified by Dil-acLDL+FITC-UEA-1 double fluorescence staining method.The human EPCs successfully identified were treated with 30 mmol/L glucose for 120 h,respectively,to make a cell model of oxidative stress damage induced by high sugar. The EPCs was divided into Astragaloside + high-glucose impaired EPCs group, Astragaloside + high-glucose impaired EPCs+ML385 group, high-glucose impaired EPCs group and normal EPCs group (5 mmol/L glucose intervention for 120 h). The levels of reactive oxygen species (ROS) were detected by flow cytometry, and the expression of oxidative stress-related proteins and Nrf2, Keap1 and NFE2L2/ARE-dependent signal pathway protein were detected by Western blotting. Result: Compared with the normal EPCs group, the intracellular ROS level was significantly increased in high-glucose impaired EPCs group. And the activities of GSH-Px, CAT and SOD were significantly decreased in high-glucose impaired EPCs group, whule the activities of LDH were significantly increased. The expressions of Nox2 and Keap were significantly increased in high-glucose impaired EPCs group, while the expressions of Nrf2, HMOX1, xCT and GPX4 were significantly decreased, with significant differences (P<0.05). Compared with high-glucose impaired EPCs group, the intracellular ROS level decreased in Astragaloside + high-glucose impaired EPCs group. And the GSH-PX, CAT and SOD activity increased in Astragaloside + high-glucose impaired EPCs group, while the LDH activity decreased. The Nox2 and Keap1 expression decreased in Astragaloside + high-glucose impaired EPCs group, while Nrf2, HMOX1, xCT and GPX4 expression increased, with significant differe

关 键 词：内皮祖细胞 氧化应激 黄芪甲苷 高糖 活性氧 

分 类 号：R285.5[医药卫生—中药学]