摘要
目的 研究肿瘤坏死因子 (TNFα)对血管内皮细胞通透性的影响及其机理。方法 采用生物素标记白蛋白的酶联免疫吸附法测定人脐静脉内皮细胞 (HUVEC)单细胞层的通透性 ;免疫荧光、激光共聚焦显微镜和蛋白免疫印迹等方法测定血管内皮黏附因子 (VEcadherin)的分布 ;功能性激酶试验测定有丝分裂原激活蛋白激酶 (MAPK)的活性 ,SB2 0 2 190 (P38抑制剂 )与PD980 5 9(ERK抑制剂 )复合物用于其活性的抑制实验。结果 与对照组比较 ,TNFα明显增加血管内皮细胞的通透性、降低细胞膜VEcadherin的表达 ,诱导P38和ERKMAPK的活性 (P <0 .0 5 )。有趣的是SB2 0 2 190明显降低了TNFα对血管内皮通透性和细胞膜VEcadherin表达的作用 ,而PD980 5 9不能阻止TNFα产生的上述作用。结论 TNFα增加血管内皮细胞的通透性是通过激活P38MAPK信号系统进而抑制VEcadherin在细胞膜的表达和重新分布。
Objective To study the influenc e of tumor necrosis factor alpha (TNFα) on endothelial permeability and its m echanism. Methods Human umbilical vein endothelial cell (HU VEC) monolayer permeability was measured by enzyme-linked immunosorbent assay f or biotin-labeled albumin. Immunofluorescence, laser confocal microscopy, and W estern immunoblotting were used to assess vascular endothelial (VE) cadherin dis tribution. Mitogen-activated protein kinase (MAPK) activity was determined by u sing functional kinase assay and was inhibited with the compounds SB202190 (P38 inhibitor) and PD98059 (ERK inhibitor). Results TNFα s ignificantly increased endothelial permeability and reduced the expression of me mbrane-associated VE cadherin. Furthermore, TNFα activated P38 MAPK and ER K MAPK compared with controls (P<0.05). Interestingly, SB202190 significantl y reduced the effect of TNFα on endothelial permeability and cell-surface VE cadherin expression, but PD98059 did not. Conclusion TNF α increases endothelial permeability by activating P38 MAPK and then inhibi ting the expression of VE cadherin and its redistribution on HUVEC.
出处
《西安交通大学学报（医学版）》
CAS
CSCD
北大核心
2004年第6期538-541,548,共5页
Journal of Xi’an Jiaotong University（Medical Sciences）
