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目的:探讨冠心病患者循环内皮祖细胞(EPCs)数量及功能变化与血管内皮舒张功能的关系。方法:将58例患者分为对照组(20例)、稳定性心绞痛组(11例)、不稳定性心绞痛组(27例)。采用高分辨率二维超声检测肱动脉血流介导的内皮依赖性血管舒张功能(FMD)及硝酸甘油介导的非内皮依赖性血管舒张功能(NMD);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,培养7天后贴壁细胞进行细胞化学分析,激光共聚焦显微镜鉴定异硫氰酸荧光素标记荆豆凝集素I和DiI标记的乙酰化低密度脂蛋白双染色阳性细胞为正在分化的EPCs,采用二苯基四氮唑嗅盐比色法、改良的Boyden小室和黏附能力测定实验观察内皮祖细胞的增殖能力、迁移能力和黏附能力。结果:①稳定性心绞痛组与不稳定性心绞痛组的FMD均明显低于对照组,有显著性差异(P<0．05～0．01),而稳定性心绞痛组的FMD较不稳定性心绞痛组也降低,有显著性差异(P<0．05);3组间NMD差异无统计学意义(P>0．05)。②稳定性心绞痛组、不稳定性心绞痛组较对照组循环EPCs数量明显减少,且黏附、迁移及增殖能力也明显下降,均有极显著性差异(P<0．01);不稳定性心绞痛组较稳定性心绞痛组EPCs数量及迁移能力无差异(P>0．05),但黏附和增殖能力降低(P<0．05)。③直线相关性分析发现不稳定性心绞痛组EPCs的数量及功能均与FMD呈正相关(P<0．05),而稳定性心绞痛组仅EPCs黏附功能与FMD呈正相关(P<0．05)。结论:冠状动脉EPCs数量及功能下降与血管内皮依赖性舒张功能障碍一致,提示当冠心病患者血管内皮功能受损而又缺乏足够有效的EPCs时,可能影响冠心病的病情程度及临床表现。
Objective: To investigate the relationship between the number and function of circulating endothelial progenitor cells (EPCs) and the function of vascular endothelial relaxation in patients with coronary heart disease. Methods: 58 patients were divided into control group (20 cases), stable angina group (11 cases) and unstable angina group (27 cases). Brachial artery-mediated endothelium-dependent vasodilation (FMD) and nitroglycerin-mediated non-endothelium-dependent vasodilatation (NMD) were measured by high resolution two-dimensional echocardiography. Peripheral blood was obtained by density gradient centrifugation Mononuclear cells were seeded on human fibronectin coated plates and cultured for 7 days. The adherent cells were subjected to cytochemical analysis. Confocal laser scanning microscopy was used to identify fluorescein isothiocyanate labeled Vitexin L and DiI labeled acetyl EPCs were differentiated into EPCs. The proliferation, migration and adhesion of endothelial progenitor cells were evaluated by diphenyltetrazolium bromide salt assay, modified Boyden chamber and adhesion assay . Results: ① The FMD of stable angina group and unstable angina group were significantly lower than that of the control group (P <0.05 ~ 0.01), while FMD of stable angina pectoris group was lower than that of unstable angina pectoris group There was also a significant difference between the three groups (P <0.05). There was no significant difference in NMD between the three groups (P> 0.05). ② The numbers of circulating EPCs in stable angina group and unstable angina group were significantly decreased compared with those in control group, and the adhesion, migration and proliferation ability also significantly decreased (P <0.01); unstable angina group The number of EPCs and migration ability in the patients with stable angina pectoris had no difference (P> 0.05), but the adhesion and proliferation ability decreased (P <0.05). (3) The linear correlation analysis showed that the number and function of EPCs in unstable angina group were positively correlated with FMD (P <0.05), while only the adhesion function of EPCs in stable angina pectoris group was positively correlated with FMD (P <0.05). Conclusions: The number and function decline of EPCs in coronary arteries are consistent with endothelium-dependent dilation dysfunction, which suggests that coronary artery disease may affect the severity and clinical manifestations of coronary heart disease when it has impaired vascular endothelial function and lack of enough effective EPCs.