系统性硬皮病患者外周血内皮祖细胞数量和功能的研究

目的：测定系统性硬皮病患者外周血中的内皮祖细胞（endothelial progenitors cells，EPCs）数量和增殖、迁移、黏附功能，探讨EPCs在其发病中的作用。方法：系统性硬皮病患者和健康者各20例。密度梯度离心法分离外周血单个核细胞并培养，流式细胞仪检测CD133^＋、CD34^＋或CD133^＋、VEGFR2^＋双荧光标记的EPCs数。MTT比色法、Millicell小室和黏附能力测定实验分别检测EPCs的增殖、迁移和黏附能力。结果：系统性硬皮病患者外周血EPCs数为（0．10±0．09）％，健康对照组为（0．49±0．09）％，P〈0．01，系统性硬皮病患者外周血EPCs数较健康者减少。实验组外周血EPCs增殖、迁移和黏附能力分别为（13．01±4．33）％、（7．20±3．67）个／视野和（15．20±4．40）个／视野，健康对照组分别为（23．28±4．34）％、（14．15±2．98）个／视野和（24．80±3．58）个／视野，两组比较，P值均〈0．01，实验组EPCs增殖、迁移和黏附能力均较对照组有所下降。结论：系统性硬皮病患者外周血EPCs数量减少，增殖、迁移和黏附功能降低，可能参与了系统性硬皮病的发病。

STUDY OF THE CHANGES OF NUMBER AND ACTIVITY OF ENDOTHELIAL PROGENITOR CELLS IN PERIPHERAL BLOOD FROM SYSTEMIC SCLERODERMA PATIENTS

Objective：To study the changes of number, proliferation,migration and adhesion function of endothelial progenitors cells （EPCs） in peripheral blood from systemic scleroderma patients, and try to explore their roles in the pathogenesis of systemic scleroderma. Methods：Twenty patients were included in this study together with 20 healthy people as controls. By ficoll density gradient centrifugation, mononuclear ceils were isolated from peripheral blood. Cells with double markers of CD133^＋, CD34^＋ or CD133^＋, VEGFR2^＋, were identified through flow cytometry. MTT trial was utilized to evaluate proliferation of EPCs. Migration and adhesion function of EPCs were assessed by modified millicell chamber assay and adherent activity assay. Results：The counting of EPCs in experimental and control group were （0.10±0.09）% and （0.49±0.09）%,the difference was statistically significant.Cell growth rates of experimental and control group were （13.01 ±4.33） % and （23.28 ±4.34）% respectively.The ability of EPCs migration and adhesion in experimental group were（7.20±3.67）/HPF and （15.20±4.40）HPF, in contrast to（14.15±2.98）/HPF and（24.80 ±3.58） / HPF in control group. Therefore, function of EPCs including migration and adhesion activity was reduced. Conclusion：The number and function of EPCs are identified to be decreased in systemic scleroderma patients, which may take part in the pathogenesis of systemic scleroderma.