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目的观察不稳定型心绞痛(UA)患者外周循环及心脏局部内皮祖细胞(EPCs)数量及功能的改变。方法27例UA患者均经冠状动脉造影(CAG)证实有明显冠状动脉狭窄,20例对照组患者经临床检查和CAG排除冠心病。密度梯度离心法从外周循环及冠状窦获取单个核细胞,将其接种在人纤维连接蛋白包被培养板,培养14 d后贴壁细胞进行细胞化学分析。激光共聚焦显微镜鉴定FITC-UEA-I和DiI-acLDL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EPCs的增殖能力、迁移能力和黏附能力。结果①与对照组比较,UA患者外周循环及心脏局部EPCs数量均明显减少,EPCs的黏附、迁移及增殖能力也显著受损(P<0.01)。②UA患者冠状窦EPCs的数量增加,黏附和增殖能力较外周循环有所下降(P<0.05),迁移能力无明显差别。结论UA患者心脏局部EPCs的数量下降和功能降低,提示冠状动脉内皮损伤后修复能力低下,可能与病情加重有关。
Objective To observe the changes of the number and function of peripheral circulation and cardiac endothelial progenitor cells (EPCs) in patients with unstable angina pectoris (UA). Methods Twenty-seven patients with UA were confirmed coronary artery stenosis by coronary angiography (CAG). The patients in the control group were excluded from coronary heart disease by clinical examination and CAG. The mononuclear cells were obtained from the peripheral circulation and the coronary sinus by density gradient centrifugation. The cells were inoculated on human fibronectin coated culture plates, and the adherent cells were cultured for cytochemical analysis after 14 days. Confocal laser scanning microscopy identified FITC-UEA-I and DiI-acLDL double staining positive cells as differentiated EPCs. The proliferation, migration and adhesion of EPCs were observed by diphenyltetrazolium chloride salt assay, modified Boyden chamber and adhesion assay. Results ① Compared with the control group, the numbers of peripheral circulation and cardiac EPCs in UA patients were significantly decreased, and the adhesion, migration and proliferation of EPCs were also significantly impaired (P <0.01). ② The number of EPCs in coronary sinus of UA patients increased, the adhesion and proliferation ability decreased compared with that in peripheral circulation (P <0.05), and there was no significant difference in migration ability. Conclusions The number of EPCs in the heart of UA patients is decreased and the function is decreased. It suggests that the repair ability of endothelial cells after coronary artery injury may be related to the worsening of the disease.