摘要
目的观察黄芪甲苷对体外培养的人外周血内皮祖细胞(EPCs)数量及增殖、迁移、黏附功能、血管生成能力的影响。方法密度梯度离心法获取人外周血单个核细胞,FITC标记荆豆凝集素Ⅰ(FITC-UEA-I)、DiI标记的乙酰化低密度脂蛋白(DiI-acLDL)荧光双染鉴定,流式细胞仪检测细胞表面标志。单个核细胞培养4d后进行实验分组,分为对照组和黄芪甲苷干预组。干预组加入不同浓度的黄芪甲苷(分别为10μg/mL、20μg/mL、40μg/mL)培养72h。分别采用四氮唑溴盐比色法、改良的Boyden小室和黏附能力测定来观察EPCs的增殖、迁移和黏附能力;体外血管生成试剂盒来观察EPCs体外血管生成能力。结果与正常对照组比较,黄芪甲苷组EPCs数量增加,EPCs的黏附、迁移、增殖和体外血管形成能力增强,且黄芪甲苷20μg/mL时作用最显著。结论黄芪甲苷可增加体外培养的EPCs数量,改善EPCs增殖、迁移、黏附和体外血管生成能力等生物学功能。
Objective To investigate the effects of astragaloside Ⅳ on vasculogenesis of peripheral blood-derived endothelial progenitor cells（EPCs） in vitro.Methods Peripheral blood monocytes（PBMCs） were isolated by ficoll density gradient centrifugation from peripheral blood and cultured.The laser scanning confocal microscope and flow cytometry（FCM） were used to determine the EPCs in human peripheral blood.The EPCs were incubated with control serum and different concentration astragaloside Ⅳ-containing cultivate liquid（10 μg/mL,20 μg/mL,40 μg/mL） for 72 hours.The activity of EPCs was observed under reverse microscope.MTT was used to determine the optical density（OD） value of EPCs.Numbers of EPCs,and the ability of cell adhesion was assessed by cell counting and adhesion assay.In vitro vasculogenesis activity was assayed by in vitro vasculogenesis kit.Results Compared with control group,the number of cultured EPCs were remarkably increased,and the proliferation,migration,adhesive function and in vitro vasculogenesis capacity of EPCs were remarkably improved in astragaloside Ⅳ groups.In addition,medium dosage astragaloside Ⅳ（20 μg/mL） had the most prominent effects.Conclusion Astragaloside Ⅳ could increase the number of cultured EPCs as well as promotes its proliferation,migration,adhesive function and in vitro vasculogenesis capacity of peripheral blood-derived EPCs.
出处
《中西医结合心脑血管病杂志》
2012年第10期1214-1216,共3页
Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease
关键词
黄芪甲苷
内皮祖细胞
血管生成能力
astragaloside Ⅳ
endothelial progenitor cells
vasculogenesis
