晚期糖基化终末产物刺激下内皮细胞活性氧的变化及来源分析  

Increased Reactive Oxygen Species in Endothelial Cells Stimulated by Advanced Glycation End Products Mediated by NADPH Oxidase

作　　者：于世勇[1] 黄岚[1] 宋明宝[1] 赵晓辉[1] 陈剑飞[1] 崔斌[1] 

机构地区：[1]中国人民解放军第三军医大学新桥医院全军心血管内科研究所,重庆市400037

出　　处：《中国动脉硬化杂志》2008年第11期857-860,共4页Chinese Journal of Arteriosclerosis

基　　金：国家自然科学基金(30470729);全军医学科学技术研究"十一五"计划课题(06J013)

摘　　要：目的探讨晚期糖基化终末产物刺激下内皮细胞活性氧的变化及来源。方法培养人脐静脉内皮细胞,观察不同浓度和不同作用时间晚期糖基化终末产物刺激下人脐静脉内皮细胞内活性氧的生成情况,进而分别应用线粒体呼吸链酶复合体、黄嘌呤氧化酶、一氧化氮合酶及NADPH氧化酶抑制剂,观察抑制相应氧化酶后细胞内活性氧的变化,分析各种氧化酶作用大小。结果晚期糖基化终末产物刺激后,内皮细胞内活性氧明显增加,并且随晚期糖基化终末产物浓度和孵育时间的增加而增加,NADPH氧化酶抑制剂二亚苯基碘几乎完全抑制了晚期糖基化终末产物刺激下细胞内活性氧的生成,线粒体呼吸链酶复合体I抑制剂鱼藤酮、线粒体呼吸链酶复合体Ⅱ抑制剂噻吩甲酰三氟丙酮、线粒体呼吸链酶复合体Ⅲ抑制剂抗霉素A、黄嘌呤氧化酶抑制剂别嘌醇对活性氧生成无明显影响,而一氧化氮合酶抑制剂左旋硝基精氨酸甲酯反而引起细胞内活性氧的轻度增加。结论NADPH氧化酶是晚期糖基化终末产物刺激下内皮细胞活性氧生成的主要来源,可能是晚期糖基化终末产物启动和加重动脉粥样硬化病理生理过程中的重要防治靶点。Aim Advanced glycation end products （ AGE ） can induce the intracellular generation of reactive oxygen species （ ROS）and cause the dysfunction of endothelial cells. This may accelerate development of vascular atherosclerosis. The source of intracellular ROS in endothelial cells stimulated by AGE was investigated. Methods Human umbilical vein endothelial cells （HUVEC ） were cultured and incubated with different concentration of AGE for different time. Intracellular ROS was measured with 2 ＇, 7 ＇ -dichlorodihydrofluorescein diacetate （ DCF-DA ）. rotenone, thenoyltrifluoroacetone （ TTFA ） and antimycin A as selective inhibitors of mitochondrial complex Ⅰ, Ⅱ and Ⅲ were used. allopurinol as an inhibitor of xanthine oxidase, Nω-Nitro-L-arginine methyl ester （ L-NAME ） as an inhibitor of nitric oxide synthase, and diphenylene iodonium （DPI） as an inhibitor of NADPH oxidase were also used. The role of each oxidase in the generation of ROS was observed. Results Intracellular ROS was elevated by the stimulation of AGE. DPI almost completely inhibited the generation of ROS. No significant effect was observed in rotenone, TTFA, antimycin A or allopurinol, while L-NAME increased the level of ROS slightly. Conclusions Tthe results demonstrate that NADPH oxidase is the major source of intracellular ROS in endothelial cells, and the potential targets for the inhibition of the atherogenic signals triggered by AGE.

关 键 词：病理学与病理生理学 晚期糖基化终末产物 内皮细胞 活性氧 

分 类 号：R363[医药卫生—病理学]