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晚期糖化终产物诱导内皮细胞通透性增高

郭晓华, 黄巧冰, 陈波, 王述昀, 侯凡凡, 富宁

南方医科大学病理生理学教研室.广东,广州 510515；南方医科大学南方医院肾内科.广东,广州 510515；南方医科大学免疫学教研室.广东,广州 510515

摘要

该文探讨了晚期糖化终产物(advanrced glycation end products,AGEs)修饰蛋白对内皮细胞通透性及细胞骨架肌动蛋白的形态学影响,以及特异的AGEs受体(receptors for AGEs,RAGE)、氧化应激和p38 MAPK通路在此病理过程中的作用。用不同浓度的AGEs修饰人血清白蛋白(AGE--HSA)与人脐静脉内皮细胞株ECV304在体外共同培养不同时间,并设立对照组进行比较,采用TRITC荧光标记白蛋白漏出法测定单层内皮细胞的通透系数{sl Pa}值,荧光染色法示细胞骨架的形态学改变。与对照组相比,AGE--HSA以时间和剂量依赖的方式引起单层内皮细胞通透性的升高及细胞骨架肌动蛋白F--actin形态的改变;可溶性RAGE的抗体(anti--RAGE IgG)、NADPH氧化酶抑制剂(apocynin)及p38抑制剂SB203580均可减轻AGEs对内皮细胞屏障功能和形态的影响。结果提示,AGEs修饰蛋白对单层内皮细胞通透性及骨架重排的作用可能通过与内皮细胞上的RAGE结合,引起细胞内的氧化应激,并激活p38 MAPK通路所介导。

关键词： 晚期糖化终产物; 内皮细胞通透性; 细胞骨架; 晚期糖化终产物受体; NADPH氧化酶; p38 MAPK通路

Mechanism of advanced glycation end products-induced hyperpermeability in endothelial cells

Guo Xiaohua, Huang Qiaobing, Chen Bo, Wang Shuyun, Hou Fanfan, Fu Ning

Department of pathophysiology, The Southern Medical University.Guangzhou 510515,Guangdong；China；Department of Immunology, The Southern Medical University.Guangzhou 510515,Guangdong

Abstract

The purpose of the present study was to investigate the effects of advanced glycation end products (AGEs) modified protein on the permeability of endothelium monolayers and morphological changes of actin cytoskeleton. The roles of receptor for AGEs (RAGE), oxidant stress and the activation of p38 MAPK pathway in this pathological procedure were elucidated. Human umbilical vein endothelial cells (HUVECs)-derived cell line (ECV304) were incubated with AGEs modified human serum albumin (AGE-HSA) in concentrations of 12.5, 25, 50, and 100 mg/ml respectively, for 2, 4, 8, 12 and 24 hours. As control, HSA of the same concentration was administered to cells. Then TRITC-albumin was added to evaluate Pa value that reflects the permeability of endothelial monolayer. Furthermore, to visualize the morphological changes of actin cytoskeleton, the treated cells were incubated with rhodamine-phalloidin to stain F-actin. The results showed that the trans-endothelial membrane flux of albumin was significantly increased in a concentration- and time-dependent manner upon the stimulation of AGE-HSA, accompanying with actin reorganization. The blockage of AGE and RAGE binding with anti-RAGE IgG and the pharmacological inhibition of NADPH oxidase or p38 MAP kinase greatly attenuated the AGE-induced hyperpermeability response, respectively. These results indicate that RAGE, NADPH oxidase and p38 MAPK are possibly involved in the mediation of AGEs-induced barrier dysfunction and actin cytoskeleton reorganization in endothelial cells.

Key words: advanced glycation end products (AGEs);endothelial cells monolayers permeability;actin cytoskeleton;RAGE;NADPH oxidase;p38 MAPK

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引用本文：

郭晓华, 黄巧冰, 陈波, 王述昀, 侯凡凡, 富宁. 晚期糖化终产物诱导内皮细胞通透性增高[J]. 生理学报 2005; 57 (2): .

Guo Xiaohua, Huang Qiaobing, Chen Bo, Wang Shuyun, Hou Fanfan, Fu Ning. Mechanism of advanced glycation end products-induced hyperpermeability in endothelial cells. Acta Physiol Sin 2005; 57 (2): (in Chinese with English abstract).