摘要
目的:观察人肿瘤坏死因子-α(TNF-α)对人血管内皮细胞(EA.hy926)单层通透性的影响并初步探讨其作用机制。方法:测定异硫氰酸荧光素(FITC)标记的葡聚糖透过Transwell小室的荧光强度,以表示EA.hy926细胞单细胞层的通透性大小;激光共聚焦显微镜观察细胞骨架肌动蛋白(F-actin)和血管内皮钙黏蛋白(VE-cadherin)的形态分布;蛋白免疫印迹检测钙黏蛋白的表达。结果:与对照组比较,TNF-α使EA.hy926内皮细胞的通透性明显增加(P<0.05),诱导肌动蛋白重新分布及应力纤维形成,并使钙黏蛋白排列紊乱、断裂、细胞间裂隙形成增多。免疫印迹检测表明TNF-α减少钙黏蛋白表达呈剂量和时间依赖性。结论:TNF-α诱导内皮细胞通透性增高,其机制可能与其破坏内皮细胞屏障功能的完整性有关。
Objective To study the effect of human tumor necrosis factor alpha （TNF-α） on human vascular endothelial cell monolayer membrane permeability and its mechanism. Methods Human vascular endothelial cell （EA.hy926） monolayer permeability was measured by detecting fluorescence intensity of dextran labeled with fluorescein isothiocyanate （FITC） in the lower compartment of Transwell. Immunofluorescence and laser confocal microscopy were used to assess vascular endothelial actin cytoskeleton and VE-cadherin distribution. Western blotting was used to assess VE-cadherin expression. Results Compared with control group, TNF-ct significantly increased endothelial permeability （P〈0.05） and induced F-actin redistribution and stress fiber formation with VE-cadherin derangement and break up. Gaps between ECs increased obviously. Furthermore, Western blotting showed that TNF-α reduced VE-cadherin expression in a dose- and time-dependent manner. Conclusions TNF-α increased endothelial cell permeability by damaging integrity of endothelial barrier function.
出处
《外科理论与实践》
2009年第3期323-327,共5页
Journal of Surgery Concepts & Practice
基金
上海市重点学科建设项目资助(S30204)
作者简介
通讯作者：汤耀卿．E-mail：yaoqt@live．cn
