目的 探讨系统性硬皮病(Systemic scleroderma,SSc)患者外周血内皮祖细胞(endothelial progenitor cells,EPCs) 数量和分化功能的变化.方法 选取女性 SSc 患者及健康志愿者各 20 例,抽取外周血 20 ml,密度梯度离心法分离单个核细 胞,以 CD133/CD34、CD133/VEGFR2 双荧光标记鉴定细胞,CD34/CD133/VEGFR2 三荧光标记流式检测 EPCs 数量;分别以 PE 标记抗人 CD14,FITC 标记抗人 CD64,PerCP/Cy5.5 标记抗人 VEGFR2,流式细胞仪检测单阳性细胞率,进而比较分化功能.结果 SSc 患者外周血 EPCs 表达阳性率(3.18±1.97)%,较健康对照的(20.56±4.37)%明显下降,差异有统计学意义(P ＜ 0.01);培养 7 天后,外周血单个核细胞表达单核细胞表面标志 CD14+、CD64+细胞百分率(32.49±5.41)%、(30.57±4.83)%,与对照组的(14.76±2.37)%、(15.39±2.09)%比较明显增加,差异均有统计学意义(均 P ＜0.01),而内皮细胞表面标记 KDR+细胞百分率(68.38±4.65)%与对照组的(89.81±5.13)%比较明显减低,差异有统计学意义(P ＜0.01).结论 SSc 患 者循环内皮祖细胞数量减少,分化功能降低.%Objective To investigate the number and differentiation function of circulating endothelial progenitor cells (EPCs) of patients with systemic scleroderma ( SSc ) . Methods Twenty female patients with SSc and 20 healthy female volunteers were enrolled for the study. Density gradient centrifugation was used to isolate mononuclear cells (MNCs) from peripheral blood 20 ml,and EPCs were identified by analyzing both the fluorescence expression of CD 34 and CD133 ,or both CD133 and VEGFR2. The number of EPCs wasflow-eytometrically determined. We marked the cells with PE-CD14,FITC-CD64,and KDR-VEGFR2 respectively, and confirmed the single-positive cells rates by FCM ,then compared the differentiation of functions between the two groups . Results Compared with the healthy control, the number of EPCs significantly decreased in the pa - tients of experimental group [(3.18% ±1.97)% vs(20.56% ± 4. 37)% ,P < 0. 01 ], and the expression of CD14+ and CD64+ cells in the patients with SSc increased significantly [ (32.49 ± 5.41 )% vs(14.76 ± 2. 37)% , (30. 57 ± 4. 83)% vs(15. 39 ± 2.09)% , P < 0.01 ] ,but the expression of KDR+ cells in the patients with SSc decreased significantly [ (68. 38 ± 4. 65 )% vs(89. 81 ±5. 13)%,P<0.01]. Conclusion The number of the circulating EPCs greatly decreased and the function of differentiation of the cells was impaired in the patients with SSc.
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