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

作者：陈 艳1，高入春1，广可颜2，孙 忞2，彭光霖2，瞿文静1，贺有缘1，赵思佳1，邹晓玲1，熊 武2

单位：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

引用：引用：陈艳，高入春，广可颜，孙忞，彭光霖，瞿文静，贺有缘，赵思佳，邹晓玲，熊武.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[J].中医药导报,2025,31(1):8-12,19.

DOI：10.13862/j.cn43-1446/r.2025.01.002

PDF： 下载PDF

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。

关键词：内皮祖细胞；氧化应激；黄芪甲苷；高糖；活性氧

Abstract：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 differences (P<0.05). Conclusion: High-glucose can induce oxidative stress injury of EPCs, and Astragaloside can play an anti-oxidative stress role by mediating NFE2L2 (Nrf2) /ARE signaling pathway, protecting and improving endothelial progenitor cells damaged by high glucose.

Key words：endothelial progenitor cells; AstragalosideIV; oxidative stress; high-glucose ; active oxygen

发布时间：2025-11-28

点击量：58