本研究旨在观察阿司匹林对人脐血晚期内皮祖细胞(endothelial progenitor cells,EPC)功能的影响.采用密度梯度离心法及免疫磁珠分选法从脐血获得CD34+的单个核细胞,将其接种在人纤维连接蛋白包被的培养板上,培养2周后收集贴壁细胞,通过流式细胞术、免疫荧光、RT-PCR、DiI-Ac-LDL的摄取及体外血管形成能力等方法对其进行鉴定.用不同浓度阿司匹林(终浓度分别为0.1,1,10,100,1000,10000 μmol/L)处理EPC 24 h,然后分别采用CCK-8比色法、Transwell小室、黏附能力测定实验来观察EPC的增殖能力、迁移能力和黏附能力的变化.结果表明:低浓度的阿司匹林(0.1-1000 μmol/L)对脐血晚期EPC的增殖无明显影响,但可增强晚期EPC的黏附及迁移能力,而高浓度的阿司匹林(10000 μmol/L)显著抑制晚期EPC的增殖和迁移,但对其黏附能力无明显影响.结论:低浓度的阿司匹林可增强晚期EPC的黏附及迁移能力,而高浓度的阿司匹林抑制晚期EPC的增殖、黏附及迁移能力,从而抑制内皮细胞的生成.%This study was aimed to investigate whether aspirin has effect on function of late endothelial progemtor cells (EPC).Cord blood CD34 + cells were purified using the ficoll density gradient centrifugation and human CD34 positive selection kit,then the cells were inoculated on fibronectin-coated culture plate.After culture for 2 weeks,adherent cells were identified as EPC by flow cytometry,immunofluorescence,RT-PCR,uptake of Dil-Ac-LDL and matdgel tube formation assay.EPC were treated with different concentrations of aspirin (0.1,1,10,100,1000,10000 μmol/L) for 24 h,then the proliferation,adhesion and migration ability of these cells were analyzed by CCK-8 assay and transwell methods.The results indicated that the low concentrations of aspirin (0.1 and 1000 μmol/L) promoted late EPC adhesive and migratory capacity,but no obvious effect on proliferation of late EPC were observed.On the other hand,the high concentrations of aspirin (10000 μnol/L) inhibited proliferation and migratory capacity of EPC,but had no obvious effect on adhesive ability of EPC.It is concluded that low concentration of aspirin promotes migration and adhesion of late EPC,while the high concentration of aspirin decreases EPC proliferation and migratory capacity of EPC.
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