肿瘤坏死因子-α对血管内皮细胞通透性及细胞骨架和紧密连接形态变化的诱导作用  

A study on increased permeability and morphological changes in actin cytoskeleton and tight junction of vascular endothelial cells induced by tumor necrosis factor-α

作　　者：郑运江[1] 汤耀卿[1] 刘伟[1] 毛恩强[1] 李磊[1] 武均[1] 张如愿[1] 张圣道[1] 

机构地区：[1]上海交通大学医学院附属瑞金医院外科ICU,200025

出　　处：《中国危重病急救医学》2009年第3期160-163,I0001,共5页Chinese Critical Care Medicine

基　　金：上海市先进学科项目支持（S30204）

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。方法在培养的EA．hy926融合成单细胞层后，加入5、10、20／zg／LTNF—α培养24h，或加入10μg／LTNF—α培养6、12、24h，测定异硫氰酸荧光素（FITC）标记的葡聚糖（Dextran）荧光强度以表示人血管内皮细胞单细胞层的通透性大小；用激光共聚焦显微镜观察内皮细胞肌动蛋白骨架[纤维状肌动蛋白（F—actin）]和紧密连接蛋白（zO一1）的形态分布；用蛋白质免疫印迹法（Western blotting）检测ZO-1的表达。结果与空白对照组比较，TNF—α能明显增加内皮细胞的通透性，诱导F—actin的重新分布以及应力纤维的形成和ZO-1的排列紊乱，数量减少，细胞间裂隙形成增多。Western blotting表明，TNF—α呈剂量和时间依赖性的方式减少ZO—1表达。结论TNF—α诱导内皮细胞通透性增高与其破坏内皮细胞屏障功能完整性有关。Objective To study the effect of human tumor necrosis factor-α （TNF-α） on permeability of human vascular endothelial cell （EA. hy926） monolayer and its mechanism. Methods 5, 10, 20μg/L TNF-α was respectively added to the cultured endothelial cell monolayer for 24 hours, or 10μg/L TNF-α for 6, 12, 24 hours. Human vascular endothelial cell （EA. hy926） monolayer permeability was measured by detecting fluorescence intensity of fluorescein isothiocyanate （FITC） labeled dextran. Immunofluorescence and laser confocal microscopy were used to assess vascular endothelial actin cytoskeleton （F-actin） and tight junction protein （ZO-1） distribution. Western blotting was used to assess ZO-1 expression. Results Compared with control group, TNF-α significantly increased endothelial permeability and induced F-actin redistribution and stress fiber formation with ZO-1 derangement. Gaps increased obviously between endothelial cells. Furthermore, Western blotting showed that TNF-α reduced ZO-1 expression in a dose- and time-dependent manner. Conclusion TNF-α increased endothelial cell permeability by damaging integrity of endothelial barrier function.

关 键 词：肿瘤坏死因子 血管内皮细胞 通透性 肌动蛋白 紧密连接 

分 类 号：R686[医药卫生—骨科学]