形态学观察晚期糖基化终末产物诱导微血管内皮细胞核因子κB入核及其机制  

A Morphological Approach to Investigate NF-κB Nuclear Translocation Induced by Advanced Glycation End Products in Microvascular Endothelial Cells

作　　者：吴丽丽[1] 王立群[1] 王达[1] 黄巧冰[1] 

机构地区：[1]南方医科大学基础医学院病理生理学教研室广东省医学休克微循环重点实验室,广东省广州市510515

出　　处：《中国动脉硬化杂志》2013年第3期198-202,共5页Chinese Journal of Arteriosclerosis

基　　金：国家自然科学基金面上项目(30771028)

摘　　要：目的观察晚期糖基化终末产物(AGE)诱导的微血管内皮细胞核因子κB核转位,探讨氧化应激和内质网应激在核因子κB核转位中可能发挥的作用。方法用AGE修饰的牛血清白蛋白(AGE-BSA)与人皮肤微血管内皮细胞(HDMEC)在体外共同培养1 h,设立对照组进行比较,用免疫荧光化学染色示核因子κB的核转位情况;应用活性氧的抑制剂谷胱甘肽(GSH)、NADPH氧化酶(NOX)抑制剂Apocynin、内皮细胞高表达的NOX亚型NOX4的siRNA和内质网应激的标志性蛋白内质网转膜蛋白激酶1α(IRE1α)的siRNA分别预处理细胞后再给予AGE-BSA刺激,观察核因子κB的核转位情况。结果与对照组相比,AGE-BSA可诱导人皮肤微血管内皮细胞核因子κB入核;应用GSH、Apocynin、NOX4 siRNA和IRE1αsiRNA预处理细胞均可抑制核因子κB的入核。结论AGE对核因子κB的移位激活可能通过细胞内的氧化应激和内质网应激途径所介导。Aim To demonstrate the activation and nuclear translocation of the nuclear factor-κB（NF-κB） induced by advanced glycation end products （AGEs） in human dermal microvascular endothelial cells （HDMECs） , and to elucidate the roles of oxidative stress and endoplasmic reticulum stress in this pathological procedure. Methods HDMECs were incubated with AGEs-modified bovine serum albumin （AGE-BSA） at concentration of 100 mg/L for 1 h. As control, BSA of the same concentration was administered to HDMECs. NF-κB nuclear translocation was observed by immunofluorescent staining. Subsequently, HDMECs were pretreated with reduced glutathione （GSH）, apocynin, a pharmacological inhibitor of NADPH oxidase （NOX）, NOX4 siRNA or inositol requiring enzyme 1α （IRE1α） siRNA, and then administrated with AGE-BSA for 1 h. Changes of NF-κB nuclear translocation was observed. Results The results demonstrated the translocation of NF-κB from the cytoplasm to the nucleus upon the stimulation of AGE-BSA. Inhibition of reactive oxygen species generation with GSH or apocynin greatly attenuated these responses. Transfection of NOX4-small interfering RNA or IRE1α-small interfering RNA in HDMECs also significantly attenuated the AGE-induced translocation of NF-κB. Conclusion Oxidative stress and endoplasmic reticulum stress are possibly involved in the mediation of AGE-induced activation of NF-κB in endothelial cells.

关 键 词：晚期糖基化终末产物 人皮肤微血管内皮细胞 核因子ΚB 氧化应激 内质网应激 

分 类 号：R363[医药卫生—病理学]