摘要
目的:观察晚期糖基化终产物(advanced glycation end products,AGEs)对内皮细胞氧化应激水平和线粒体功能的影响,以探讨糖尿病血管内皮细胞功能障碍可能的发生机制。方法:以不同质量浓度(50,100μg/mL)的AGE修饰的牛血清白蛋白(AGE-bovine serum albumin,AGE-BSA)作用于人主动脉内皮细胞(human aortic endothelial cells,HAECs)48 h,CCK-8法测定HAECs的增殖能力,黄嘌呤氧化酶法测定超氧化物歧化酶(superoxide dismutase,SOD)活力,应用DCFH-DA探针检测细胞内活性氧(reactive oxygen species,ROS)的水平;JC-1法检测线粒体膜电位(mitochondrial membrane potential,MMP),荧光素-荧光素酶法和Clark氧电极法测定细胞中ATP含量及细胞耗氧率。结果:AGE-BSA能显著性地抑制HAECs增殖,增加细胞内ROS水平,降低SOD活力,且高质量浓度作用更加明显。同时,AGE-BSA还能使MMP下降、ATP生成及耗氧减少。结论:AGE-BSA诱导HAECs产生氧化应激损伤,其机制与其造成线粒体功能紊乱相关。
Objective:To investigate whether advanced glycation end products (AGEs) can induce cell injury and mitochondrial dysfunction in human aortic endothelial cells (HAECs).Methods:HAECs were treated with increasing concentrations (50,1 00μg/mL) of AGE-bovine serum albumin (AGE-BSA) for 48 h.The proliferative inhibition of HAECs was measured by CCK-8 method.Contents of ATP and activity of superoxide dismutase (SOD) were determined by the luciferase assay and SOD kits.Reactive oxygen species (ROS) were determined by DCFH-DA staining.Mitochondrial membrane potential (MMP) was observed with JC-1 staining.Oxygen utilization was measured polarographically with a Clark oxygen electrode.Results:Compared with control group,AGE-BSA (50,100 μg/mL) significantly reduced HAECs proliferation.AGE-BSA significantly increased the levels of ROS,while decreased the activity of SOD compared to the control group.Importantly,AGE-BSA-mediated oxidative stress was followed by a collapse of mitochondrial membrane potential (MMP),the inhibition of ATP generation,and the down-regulation of oxygen utilization.Conclusion:AGE-BSA induced oxidative stress in HAECs.Moreover,AGE-BSA-induced HAECs injury was related to mitochondrial dysfunction.
出处
《江苏大学学报(医学版)》
CAS
2014年第3期185-189,共5页
Journal of Jiangsu University:Medicine Edition
基金
国家自然科学基金资助项目(81270694)
关键词
晚期糖基化终产物
氧化应激
线粒体
人主动脉内皮细胞
advanced glycation end products(AGEs)
oxidative stress
mitochondria
human aortic endothelial cells