冠心病患者循环内皮祖细胞与血管内皮功能的变化

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。

Relationship Between Endothelial Progenitor Cells and Endothelial Function in Coronary Heart Disease Patients

Objective: To study the relationship between endothelial function and the number and activity of endothelial progenitor cells (EPCs) in coronary heart disease patients. Methods:Eleven patients with stable angina(SA) ,27with unstable angina(UA) and 20 controls were studied. Flow-mediated dilation(FMD) and nitroglycerin-mediated dilation(NMD) of brachial artery were evaluated by high-resolution ultrasound. Total mononuclear cells (MNCs) were isolated from peripheral blood by Ficoll density gradient centrifugation, and plated on hu-manfibronectin coated plates. Seven days of culture, EPCs were identified as adherent cells by double staining FITC-UEA-I binding and DiI-acLDL uptake under a laser scanning confocal fluorescence microscope. EPCs proliferation and migration activity were detected with MTT assay and modified Boyden chamber assay, respectively. EPCs adhesion assay was performed by replating MNCs on the humanfibronectin coated plates,and the adherent cells were counted. Results :(1)FMD was obviously lower in both SA group and UA group than that in control group (P <0. 05 ;P <0. 01) . It was lower in UA patients than in SA patients(P <0. 05). NMD was similar in the three groups( P >0. 05). (2)The number of EPCs was significantly reduced in UA and SA patients compared with the controls(P <0. 01). In addition, EPCs proliferative, migratory and adhesive capacity were also impaired( P <0. 01) . There was no significant difference in the number and migratory activity of EPCs between UA and SA groups( P >0. 05) , but EPCs adheresive and proliferative function was reduced in UA group compared with SA group( P < 0. 05). (3)here was a positive association between FMD versus the number and proliferative, migratory and adhesive capacity of EPCs in the UA group( P < 0. 05 ). There was a positive association between mere EPCs adhesive capacity and FMD in the SA group( P <0. 05). Conclusion: The number of EPCs is decreased and EPCs dysfunction is inversely associated with impaired endothelium-de-pendent function, which suggests that endothelial injury in the deficient circulating EPCs may affect the severity of the heart disorder and the clinical presentations.