流场对细胞黏附及其后内皮细胞VE-cadherin表达的影响  

Effects of Different Flow Fields on Cell Monocyte-endothelial Adhesion and the Changes of VE-cadherin Expression and Distribution in Endothelial Cells after Cell Adhesion

作　　者：谢爱民[1] 叶旭[1] 

机构地区：[1]湖南省肿瘤医院核医学科,湖南长沙41006

出　　处：《广州医学院学报》2008年第6期5-10,共6页Academic Journal of Guangzhou Medical College

摘　　要：目的:探讨不同流场对THP-1单核细胞与ECV304内皮细胞黏附的影响及细胞黏附后内皮细胞VE-cadherin表达及分布。方法:不同流场作用单层内皮细胞及单核细胞6h,观察不同流场下单核细胞与内皮细胞黏附;用W estern蛋白印迹法检测黏附后VE-cadherin的表达,用免疫细胞化学方法在激光共聚焦显微镜下观察VE-cadherin蛋白在内皮细胞中的分布变化。结果:不同流场作用单层内皮细胞及单核细胞6 h,发现低剪切力层流组与高剪切力层流组黏附的单核细胞均数分别为59.8/视野及3/视野,低剪切力紊流组与低剪切力层流组黏附的单核细胞均数分别为125/视野及61/视野,低剪切力紊流下单核细胞黏附数高于低剪切力层流及高剪切力层流,且在低剪切力紊流组中有大量单核细胞成簇黏附于内皮细胞上;单核细胞与内皮细胞共育0、1、2、4 h后,VE-cadherin蛋白的表达量在各组细胞间无差异;0 h组VE-cadherin着色集中于内皮细胞周边即细胞与细胞之间,1h,2h,4h时VE-cadherin着色不再集中于细胞周边而是分散到整个内皮细胞膜。结论:低剪切力紊流环境下,单核细胞与内皮细胞的黏附增加且形成了较多的成簇黏附,细胞黏附后内皮细胞膜上VE-cadherin的分布随之改变。Objective ：To elucidate the effects of different flow patterns on monocyte-endothelial cell adhesion and the changes of VE-cadherin expression and distribution in endothelial cells after cell adhesion. Methods： We investigated fluid shear regulation of monocyte-endothelial cell adhesion in vitro using different kinds of flow field to simulate the in vivo hemodynamics characteristic of lesion-prone vascular regions and normal vascular regions. Human endothelial cell monolayers were flow-conditioned for 6 h before evaluation of monoeyte adhesion under three kinds of flow fields. The expression of VE-eadherin after cell adhesion was assessed using Western blotting and the distribution of VE-cadherin protein in endothelial cells examined using laser eonfocal microscopy. Results： After treatment on endothelial monolayer and monocytes for 6 hours, the mean count of adhering monocytes per visual field was 59.8 in high sheer stress laminar group, 3 in low sheer stress laminar group, 125 in low sheer disturbed group and 61 in low sheer stress laminar group. The results revealed that there were many circulating monocytes sequestrated by endothelial and a distinctive clustered cell pattern of monocyte adhesion under low shear stress disturbed flow. The expression of VE-cadherin was comparable across all groups afler co-culture of endothelial cells and monoeytes for 0, 1, 2 and 4 hours. VE-cadherin staining at EC borders was much stronger in the group without THP-1 monocytes treatment （0 hour） than those added with monocytes and cultured for 1, 2, or 4 hours. Conclusion：This study demonstrated massive monocyte-endothelial adhesion and clustered cell pattern of monocyte adhesion under low shear stress disturbed flow. Distribution of VE-eadherin was shown to be altered after moneyte-endothelial adhesion

关 键 词：流场 单核细胞 单层内皮 黏附 VE-caherin 通透性 

分 类 号：R541.4[医药卫生—心血管疾病]