颈动脉粥样硬化患者外周血内皮祖细胞部分生物学功能改变及其意义

Changes in function of endothelial progenitor cells from peripheral blood of atherosclerosis of the carotid artery patient

目的:观察颈动脉粥样硬化患者外周血内皮祖细胞(endothelial progenitor cells,EPCs)生物学功能的改变,探讨EPCs在颈动脉粥样硬化发生发展中可能作用。方法:选择颈动脉粥样硬化患者和同年龄组正常人各20例,密度梯度离心法从外周血获取单个核细胞,接种在培养板内,培养7天后对贴壁细胞进行细胞化学分析和部分生物学功能测定。激光共聚焦显微镜鉴定FITC标记荆豆凝血素Ⅰ(FITC-UEA-I)和DiI标记的乙酰化低密度脂蛋白(DiI-acLDL)双染色阳性细胞为正在分化的EPCs。胰蛋白酶消化后计数贴壁细胞观察EPCs黏附能力;改良的Boyden小室观察EPCs的迁移能力;MTT法检测EPCs增殖能力;体外血管生成实验观察EPCs体外生成血管能力。结果:颈动脉粥样硬化患者外周血EPCs黏附细胞数(35.15±6.55)较正常对照组(45.25±7.04)显著减少(P<0.01);EPCs的增殖能力(0.649±0.033)较正常对照组(0.680±0.022)显著降低(P<0.05);颈动脉粥样硬化患者外周血EPCs体外血管生成能力较正常对照组显著减弱,并且小管的复杂程度也较对照组降低。结论:颈动脉粥样硬化患者外周血EPCs的黏附、迁移、增殖和成血管能力明显受损。EPCs的生物学功能或许可作为颈动脉粥样硬化患者血管病变发生发展的一个生物学标志。

Objective： To investigate the function of endothelial progenitor cells（EPCs） from peripheral blood of atherosclerosis of the carotid artery patient and study the role of EPCs in the development of atherosclerosis of the carotid. Methods：EPCs isolated from human peripheral blood of atherosclerosis of the carotid artery patient（n= 20）and agematched control subjects（n= 20）. Cytochemical analysis was conducted after 7days culture. EPCs were characterized as Dil-acLDL/FITC-UEA-1 double positive cell detected by laser confocalmicroscopy. EPCs adhesion assay was performed by replating those on fibronectin-coated dishes,then adherent cells were counted. The migration capcity of EPCs was determined by modified Boyden chamber. And in vitro vasculogenesis activity was assayed in vitro vasculogenesis kit. ResutsThe functional activities of EPCs such as migration, adhesion,in vitro vasculogenesis activity were significantly impaired in atherosclerosis of the carotid artery patient. Conclusion： These findings suggest that atherosclerosis of the catotid artery may alter EPCs biology,which may be play an impotant role in the devolpment of angiopathy. EPCs dysfunction may be a marker of the possibility to devolope angiopathy.