[Mechanism of advanced glycation end products-induced hyperpermeability in endothelial cells].
[Mechanism of advanced glycation end products-induced hyperpermeability in endothelial cells].
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发表时间：
2005-04
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通讯作者：
Xiao-hua Guo;Qiaobing Huang;Bo Chen;Shu-yun Wang;F. Hou;Ning Fu
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作者：
Xiao-hua Guo;Qiaobing Huang;Bo Chen;Shu-yun Wang;F. Hou;Ning Fu
本研究旨在探讨晚期糖基化终产物（AGEs）修饰蛋白对内皮单层通透性和肌动蛋白细胞骨架形态变化的影响。阐明了AGEs受体（RAGE）、氧化应激和p38 MAPK通路激活在这一病理过程中的作用。将人脐静脉内皮细胞（HUVECs）来源的细胞系（ECV304）与浓度分别为12.5、25、50和100 μ g/ml的AGEs修饰的人血清白蛋白（AGE-HSA）孵育2、4、8、12和24 h，并给予相同浓度的HSA作为对照。然后加入tritc -白蛋白评价反映内皮单层通透性的Pa值。此外，为了观察肌动蛋白细胞骨架的形态学变化，处理后的细胞用罗丹明-phalloidin孵育，染色F-actin。结果表明，AGE-HSA刺激后，白蛋白跨内皮膜通量呈浓度依赖性和时间依赖性显著增加，并伴有肌动蛋白重组。阻断AGE和RAGE与抗RAGE IgG的结合，药理抑制NADPH氧化酶或p38 MAP激酶，分别可大大减轻AGE诱导的高通透性反应。这些结果表明，RAGE、NADPH氧化酶和p38 MAPK可能参与介导ages诱导的内皮细胞屏障功能障碍和肌动蛋白细胞骨架重组。
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 microg/ml respectively, for 2, 4, 8, 12 and 24 h. 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.