胰岛素对人脐血内皮祖细胞生物活性的影响

目的探讨胰岛素对人脐血内皮祖细胞增殖、迁移与黏附功能的影响。方法采用Ficoll密度梯度离心法从人脐血中分离单个核细胞,加入EGM-2培养液,接种于包被人纤连蛋白的培养皿中贴壁培养,收集48 h后的悬浮细胞重新贴壁培养至第7天,利用免疫荧光和流式细胞术鉴定。细胞随机分为对照组和不同浓度(0.1、1和10nmol/L)胰岛素处理组干预24 h,分别采用MTT比色法、Transwell法和细胞黏附实验来观察胰岛素对内皮祖细胞增殖、迁移与黏附功能的影响。结果与对照组相比,0.1、1、10nmol/L的胰岛素干预内皮祖细胞24 h能明显增强其增殖、迁移和黏附能力(P<0.05或P<0.01);其中胰岛素浓度为1nmol/L时增殖功能达到峰值,胰岛素浓度为0.1nmol/L时迁移和黏附功能达到峰值。结论生理浓度与较低浓度胰岛素能增强内皮祖细胞的增殖、迁移与黏附能力。

Effects of insulin on activities of endothelial progenitor cells from human umbilical cord blood

Objective To investigate the effects of insulin on the proliferative,migratory and adhesive activities of endothelial progenitor cells(EPCs) from human umbilical cord blood. Methods Human umbilical cord blood mononuclear cells were isolated by Ficoll density-gradient centrifugation.EGM-2 culture fluid was added,and then cells were plated on dishes coated with human fibronectin.After 48 h,the nonadherent cells were collected and replated on fibronectin-coated dishes.Seven days later,the cells were identified with immunofluorescence and flow cytometry.The cells were randomly divided into control group and insulin groups(0.1,1 and 10 nmol/L insulin) and were incubated for 24 h.The proliferation,migration and adhesion activities of EPCs were determined with MTT assay,transwell assay and adhesive assay,respectively. Results Compared with control group,the proliferative, migratory and adhesive activities of EPCs were significantly improved after being treated with 0.1,1 and 10 nmol/L insulin for 24 h(P<0.05 or P<0.01).The proliferation activity peaked at the concentration of 1 nmol/L,and the migration and adhesion activities peaked at 0.1 nmol/L ConclusionPhysiological and lower concentrations of insulin can promote the proliferative,migratory and adhesive activities of EPCs.