冠心病患者PCI术前后循环内皮祖细胞变化

Change of endothelial progenitor cells from peripheral blood in patients with coronary heart diseases before and after percutaneous coronary intervention

目的:研究冠心病患者经皮冠脉内介入治疗(percutaneous coronary intervention,PCI)术前和术后外周血中循环内皮祖细胞(endothelial progenitors cells,EPCs)形态、数量的变化。方法:选择冠心病患者65例和对照组30例,分别在PCI手术前、术后即刻和术后4 d用密度梯度离心法从外周血获取单个核细胞,将其接种在人纤维连接蛋白包被培养板,用加入血管内皮生长因子和碱性成纤维细胞生长因子的RPMI-1640培养基培养细胞。用激光共聚焦显微镜初步鉴定DiL标记的乙酰化低密度脂蛋白和FITC标记的荆豆凝集素I双染色阳性细胞为EPCs,用流式细胞仪检测细胞表面抗原CD34和KDR;相差显微镜下分析细胞形态数量变化和集落形成单位的数量。结果:冠心病患者PCI术后EPCs数目较术前明显增加,术后4 d开始下降;PCI术后即刻较术前有明显增强,术后4 d有下降,但仍高于对照组,但PCI术前术后细胞集落形成单位的数量并无变化。结论:冠心病患者PCI手术可刺激EPCs数量的增加,在术后4 d逐渐回到术前状态。

Objective To investigate the culture of endothelial progenitor cells （ EPCs ） from peripheral blood in patients with coronary heart diseases （CHD） before and after percutaneous coronary intervention （PCI） , and to observe the cells shape and determine the cell number and proliferation activity. Methods Ninety-five patients were divided into a CHD group （n = 65 ） and a control group （ n = 30 ）. The mononuclear cells were isolated from peripheral blood of patients with CHD before, right after and 4 days after PCI by Ficoll-density centrifugation. The isolated cells were cultured in RPMI1640 medium supplemented with VEGF165 and bFGF. EPCs were characterized as adherent cells of double positive for DiL-acLDL uptake and FITC-UEA-I binding by direct fluorescent staining under a fluorescence microscope. The EPCs specific surface mark CD34 and KDR were assessed by fluorescence activated cell sorter analysis. The cell shapes were analysed and the number of colonyforming units （ CFU ） was counted by phase-contrast microscope. Results The number of EPCs reduced in patients with CHD before the PCI, but the cell number was significantly increased in patients with CHD after the PCI, and the number reduced in patients with CHD 4 days after the PCI. However, the number of CFUs did not change in patients before and after the PCI. Conclusion PCI can increase endothelial progenitor cells in patients after the PCI ; but 4 days after the PCI,this increase will not exist.