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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 effect of tumor necrosis factor (TNFα) on the permeability of vascular endothelial cells and its mechanism. Methods The permeability of monolayers of human umbilical vein endothelial cells (HUVECs) was measured by enzyme-linked immunosorbent assay (ELISA). The expressions of VE cadherin ); Functional kinase assays measure the activity of mitogen-activated protein kinase (MAPK), and the activity of SB2 0 2 190 (P38 inhibitor) and PD980 5 9 (ERK inhibitor) complexes were tested for their activity. Results Compared with the control group, TNFα significantly increased the permeability of vascular endothelial cells, decreased the expression of VE cadherin, and induced the activity of P38 and ERKMAPK (P <0.05). Interestingly, SB2O290 significantly reduced the effect of TNFα on vascular endothelial permeability and cell membrane VE cadherin expression, whereas PD980 5 9 did not prevent the above effects of TNFα production. Conclusion TNFα increases the permeability of vascular endothelial cells by activating the P38MAPK signaling system and thereby inhibiting the expression and redistribution of VE cadherin in the cell membrane.