冠心病患者外周血内皮祖细胞数量及生物学功能与肝细胞生长因子的关系

摘要

目的：观察冠心病患者外周血内皮祖细胞（EPCs）数量及生物学功能的变化，进一步探讨肝细胞生长因子（HGF）对其影响，为临床应用HGF提供理论依据。方法收集50例非冠心病患者（对照组）、50例冠心病患者（冠心病组；每例分为HGF干预组和非HGF干预组）外周血，应用流式细胞仪和ELISA法分别检测各组CD133+/CD34+细胞的数量和HGF水平；采用密度梯度离心法分离培养各组外周血中EPCs，通过MTT法、Transwell迁移试验、黏附能力测定试验及PI-AnnexinⅤ双重染色法来分别检测EPCs的增殖、迁移、黏附能力和凋亡水平。结果与对照组比较，冠心病组外周血中CD133+/CD34+细胞数量减少[（2.15±0.69）% vs （5.26±1.16）%，P＜0.01]，血浆中HGF浓度升高[（6.80±1.22） vs （2.62±0.83）µg/L，P＜0.01]，EPCs增殖、迁移、黏附等生物学功能减弱（P＜0.05）；HGF干预组EPCs增殖、迁移、黏附等生物学功能显著改善（P＜0.05）。各组细胞凋亡水平差异无统计学意义（P＞0.05）。结论外周血中CD133+/CD34+细胞数量和血浆中HGF水平的变化可能成为冠心病患者新的危险评估因素。%Objective To investigate the changes of the number and biological functions of endothelial progenitor cells (EPCs) in peripheral blood of patients with coronary heart disease (CHD) and determine the effect of hepatocyte growth factor (HGF) on the cell number and function, so as to provide theoretical basis for further clinical application of HGF. Methods A random controlled trial was carried out on 50 CHD patients and 50 control subjects matched in age, sex, hypertension, diabetes and hyperlipidemia. The number of EPCs (CD133+/CD34+) and the level of HGF in peripheral blood of each group were measured respectively by flow cytometry and ELISA. The proliferation, migration, apoptosis and adhering ability of EPCs were detected by MTT assay, Transwell migration experiment, PI-AnnexinⅤ double staining and adhering experiment, respectively. Results Compared with control group, the number of CD133+/CD34+cells in peripheral blood of CHD patients was significantly lower[(2.15±0.69)% vs (5.26±1.16)%, P<0.01]; the plasma level of HGF was higher [(6.80±1.22) vs (2.62±0.83)µg/L, P<0.01];and the ability of proliferating, migrating and adhering of EPCs was reduced(P<0.05). HGF significantly improved the biological function of EPCs isolated from patients with CHD(P<0.05), but had no effect on the apoptosis of EPCs(P>0.05). Conclusion The number of CD133+/CD34+cells in peripheral blood and plasma level of HGF might be used as new risk assessment factor for CHD patients.