辛伐他汀对人脐静脉血晚期内皮祖细胞数量和功能的影响

Effect of simvastatin on the quantity and function of human umbilical cord blood-derived late endothelial progenitor cells

目的:研究不同浓度辛伐他汀对体外培养的人脐静脉血晚期内皮祖细胞(EPCs)数量和功能的影响。方法:用密度梯度离心法分离人脐静脉血单个核细胞,接种于人纤维连接蛋白包被的培养瓶,在EGM-2MV培养基中诱导分化获得晚期EPCs,并通过流式细胞术及免疫荧光染色进行鉴定。取生长状态良好的第2代晚期EPCs与不同浓度辛伐他汀(10-8、10-7、10-6及10-5mol/L)培养24、48及72h后,采用CCK-8增殖能力检测、粘附能力测定及血管形成实验来观察辛伐他汀对晚期EPCs增殖、粘附及血管形成能力的影响。结果:从人脐静脉血中成功分离并培养出晚期EPCs。与对照组相比,较低浓度辛伐他汀(10-8、10-7与10-6mol/L)对晚期EPCs的增殖、粘附及血管形成有明显促进作用,以10-7mol/L组作用最强;高浓度(10-5mol/L)则抑制EPCs的上述功能。辛伐他汀促增殖作用随时间延长而增加,而粘附与体外血管形成能力则在48h作用最强。结论:辛伐他汀在较低浓度时显著增加了晚期EPCs的数量并改善其功能,高浓度则起抑制作用。

Objective：To investigate the effect of simvastatin on the quantity and function of late endothelial progenitor cells（EPCs）from human umbilical cord blood in vitro.Method：Mononuclear cells were isolated from human umbilical cord blood by density gradient centrifugation and cultured on fibronectin-coated culture flask, then the cells cultured in EGM-2MV medium differentiated into late EPCs which were identified by flow cytometry and fluorescent staining.The good growth state of late EPCs of second generation were treated with simvastatin in a series of concentrations（10-8,10-7,10-6,10-5 mol/L）for 24,48,72hand the proliferation,adhesion and tubule-formation activity of EPCs were analyzed by CCK-8reagent kit,adhesive assay and tubule-formation assay respectively.Result：Late EPCs were isolated and cultured successfully from human umbilical cord blood.Com- pared with control group,lower concentrations of simvastatin（10-8,10-7,10-6 mol/L）significantly promoted cell proliferation,adhesion and in vitro vascularization and the concentration of 10-7 mol/L was the most prominent,while higher concentration（1 0-5 mol/L）inhibited the functions above.Simvastatin increased the proliferative capacity of EPCs in a time dependent manner,but adhesion and tubule-formation activity were the most prominent at 48h.Conclusion：Simvastatin enhances the count of late EPCs and improve cell functions remarkably at lower concentrations,while higher concentration elicit inhibitory effects.