颈动脉硬化病人内皮祖细胞增殖和分化能力改变及其临床意义

目的观察颈动脉硬化病人外周血内皮祖细胞(EPCs)增殖和分化能力的改变,探讨其在颈动脉硬化发生发展中的意义。方法选择颈动脉硬化病人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7d后对贴壁细胞进行鉴定,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细胞。四氮唑溴盐比色法(MTT)测定EPCs的增殖能力,流式细胞仪检测细胞表面CD14 ,CD64 和vWF ,测定EPCs向内皮分化能力。结果颈动脉硬化病人外周血EPCs增殖能力较正常对照组显著降低(P<0.05);与正常对照组比较颈动脉硬化组EPCs早期标志物CD14 和CD64 百分比明显升高(P<0.05),而内皮细胞分化的特异性标志物vWF 显著降低(P<0.05)。结论动脉硬化组较正常组外周血EPCs增殖能力和分化为内皮细胞的能力明显减退,血管的再内皮化的过程受损,促进颈动脉硬化的发生发展。

Clinical Influence of Proliferation and differentiation Activity of Endothelial Progenitor Cells in Patient with Atherosclerosis of the Carotid Artery

Objective To investigate the proliferation and differentiation of endothelial progenitor cells(EPCs) from peripheral blood in patients(pts) with atherosclerosis of the carotid artery.Methods EPCs were isolated from human peripheral blood in pts with atherosclerosis of the carotid artery(n=20) and in age-matched control subjects(n=20).Cytochemical analysis was conducted after culture for 7 days.EPCs were characterized as that DiI-acLDL /FITC-UEA-I double positive cells were detected by laser confocalmicroscopy.EPC proliferation was performed by MTT assay and differentiation activity was measured by flow cytometry.Results The proliferation activity of EPCs was significantly impaired in pts with atherosclerosis of the carotid artery.The relative proportion of EPCs expressing CD14 and CD64 were increased and expressing the endothilial marker was decreased as compared with that in control subjects.Conclusion These findings suggested that EPCs proliferation and differentiation might be decreased in pts with atherosclerosis of the carotid artery,which might play an important role in the development of angiopathy.