阿司匹林对内皮祖细胞功能及其诱导型一氧化氮合酶的影响

目的:观察阿司匹林对外周血内皮祖细胞(endothelial progenitor cells,EPCs)数量和功能的影响。方法:采用密度梯度离心法从外周血获得单个核细胞,将其接种在人纤维连接蛋白包被的培养板上,培养7d后收集贴壁细胞,分别加入不同浓度阿司匹林(分别为1、2、5、10mmol/L)培养3、6、12、24h。激光共聚焦显微镜鉴定FITC-UEA-I和Dil-acLDL双染色阳性细胞为正在分化的EPCs,将其在倒置荧光显微镜下计数。然后分别采用MTT比色法、改良的Boyden小室、黏附能力测定实验和体外血管形成试剂盒来观察EPCs的增殖能力、迁移能力、黏附能力和体外血管形成能力,免疫印迹杂交法(Western blot)半定量测定诱导型一氧化氮合酶(iNOS)含量。结果:阿司匹林能减少外周血EPCs数量,并且EPCs数量随阿司匹林浓度和作用时间增加而减少。EPCs的增殖能力、迁移能力、黏附能力、体外血管形成能力和iNOS含量亦随阿司匹林浓度和作用时间增加而降低。结论:阿司匹林通过减少EPCs中iNOS含量,降低EPCs的增殖能力、迁移能力、黏附能力和体外血管形成能力,从而抑制内皮细胞的生成。

Effects of aspirin on endothelial progenitor cell function and its induced nitricoxide synthase

Objective:To investigate whether aspirin had influences on endothelial progenitor cells(EPCs).Methods:Total mononuclear cells(MNCs) were isolated from peripheral blood by ficoll density gradient centrifugation,and then the cells were plated on fibronectin-coated culture dishs.After 7 days cultured,attached cells were stimulated with aspirin(to make a series of final concentrations:1,2,5,10mmol/L)for 3,6,12,24h.EPCs were characterized as adherent cells double positive for DiLDL-uptake and lectin binding by direct fluorescent staining.EPCs proliferation,migration were assayed with MTT assay and modified Boyden chamber assay,respectively.EPCs adhesion assay was performed by replating those on fibronectin-coated dishes,and then adherent cells were counted.In vitro vasculogenesis activity was assayed by in vitro vasculogenesis kit.iNOS was assayed by western blot.Results:Incubation of isolated human MNCs with aspirin decreased the number of EPCs,aspirin also decreased EPCs proliferative,migratory,adhesive,in vitro vasculogenesis capacity and iNOS in a concentration and time dependent manner.Conclusion:Aspirin decrease the number,proliferative,migratory,adhesive and in vitro vasculogenesis capacity of EPCs,aspirin also decrease iNOS in EPCs.