黄芪注射液对外周血内皮祖细胞数量及功能的影响

目的观察黄芪对外周血内皮祖细胞(EPCs)数量、功能及诱导型一氧化氮合酶(iNOS)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,加入不同浓度黄芪(分别为10,20,50,100g/L)培养24h,同时,对照组与黄芪50g/L组分别培养6,12,24,48h。激光共聚焦显微镜鉴定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生成试剂盒来观察EPCs的增殖能力、迁移能力、黏附能力和体外血管形成能力,免疫印迹杂交法半定量测定iNOS含量。结果黄芪能增加外周血EPCs数量,促进EPCs的增殖能力、迁移能力、黏附能力和体外血管形成能力,并随黄芪浓度和作用时间的增加而增加,黄芪还能增加EPCs中iNOS含量。结论黄芪能增加EPCs的数量,促进EPCs的增殖能力、迁移能力、黏附能力和体外血管形成能力,这可能与其增加iNOS含量有关。

Effects of Astragalus on Number and Activity of Peripheral Endothelial Progenitor Cells

Objective To investigate the influence of astragalus on the number,activity,and inducible nitric oxide synthase(iNOS) of peripheral endothelial progenitor cells (EPCs). Methods Total mononuclear cells (MNCs) were isolated from peripheral blood by Ficoll density gradient centrifugation,and then the cells were inoculated on fibronectin-coated culture dishes. After 7 days of culturing,attached cells were stimulated with astragalus solution (in a series of final concentrations of 10,20,50 and 100 g/L)for 24 hours. Meanwhile,the attached cells in the control group and astragalus 50 g/L group were cultured for 6,12,24,48 h respectively. EPCs were identified by laser confocal microscopy and were counted under inverted fluorescence microscope. The proliferation and migration of EPCs were assayed with MTT assay and modified Boyden chamber assay,respectively. EPCs adhesion assay was performed by re-inoculation of EPCs on fibronectin-coated dishes,and then adherent cells were counted. In vitro vasculogenesis activity of EPCs was assayed by in vitro vasculogenesis kit. iNOS was assayed by western blot. Results Incubation of isolated human MNCs with astragalus increased the number of EPCs,as well as their proliferative,migratory,adhesive and invitro vasculogenesis capacity in a concentration and time dependent manner. Astragalus also increased iNOS content of EPCs. Conclusion Astragalus increases the number of EPCs,and enhances the proliferative,migratory,adhesive and in vitro vasculogenesis capacity of EPCs. Its mechanism is related to the increase of iNOS by astragalus.