非对称性二甲基精氨酸对肉皮生长晕细胞生物学功能的影响

目的 观察非对称性二甲基精氦酸（ADMA）对内皮生长晕细胞（EOC）及其生物学功能的影响。方法分离脐血中单个核细胞，采用贴壁培养法培养EOC，以免疫细胞化学染色及荧光染色法鉴定其内皮细胞特性。以10μmol／LADMA及配制ADMA的溶剂（含0.5％DMSO的EBM-2培养基），分别与第二代细胞孵育72h，检测并对比细胞的凋亡率、增殖能力及血管生成能力。结果免疫细胞染色后可见细胞表面Ⅷ因子相关抗原、CD34以及FIk-1均阳性表达，Dil-ac—LDL和FITC—UEA—Ⅰ荧光染色后可见双染色阳性细胞均为正在分化的EOC。ADMA组细胞凋亡率[（5.96±0．12）％]显著高于正常组[（1．48±0．03）％]（P〈0.01），而细胞增值活性及血管生成活性（0．27、0.02、8．25、0.96）均明显弱于正常组（8.25、0．96、16.25、1.71）（P〈0．05或0.01）。结论ADMA能促进EOC凋亡，抑制其增值活性及血管生成活性，其对EOC细胞功能的损伤作用可能是通过氧化应激机制实现的。

The effects of asymmetric dimethylarginine on the biological functions of endothelial outgrowth cells

Objective To investigate the effects of asymmetric dimethylarginine （ADMA） on the biological functions of endothelial outgrowth ceils （EOC）. Methods The mononuclear cells were harvested from umbilical cord blood by density gradient centrifugation, were then induced into EOC and expanded in vitro by adherent culturing method. The endothelial characteristics of EOC were identified by immunostaining and fluorescent staining. Second generation of EOCs were treated with 10 μmol/L ADMA for 72h, and in control group cells were treated with blank solution （EBM-2 medium containing 0.5% DMSO）. EOC cells were harvested and the apoptosis rate, proliferation and vasculogenesis activity were measured by flow cytometry, MTT assay and Matrix gel vasculogenesis assay respectively. Results Compared with control group, the apoptosis rate and inhibition of proliferation and vasculogenesis activity of EOCs treated with ADMA were significantly increased. Conclusion ADMA can cause damage of the biological function of EOCs.