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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 study the effects of different concentrations of simvastatin on the number and function of endothelial progenitor cells (EPCs) in human umbilical cord blood in vitro. Methods: Human umbilical vein blood mononuclear cells were separated by density gradient centrifugation, inoculated into human fibronectin coated culture flasks, and induced to differentiate into advanced EPCs in EGM-2MV medium. Flow cytometry and immunofluorescence Dye identification. The second generation of advanced EPCs with good growth condition were cultured with different concentrations of simvastatin (10-8, 10-7, 10-6 and 10-5mol / L) for 24, 48 and 72 hours, then the proliferation of CCK-8 , Adhesion assay and angiogenesis assay were used to observe the effect of simvastatin on proliferation, adhesion and angiogenesis in advanced EPCs. Results: The advanced EPCs were successfully isolated and cultured from human umbilical vein blood. Lower concentrations of simvastatin (10-8, 10-7 and 10-6 mol / L) significantly promoted the proliferation, adhesion and angiogenesis of advanced EPCs compared with the control group. 10-7mol / L group The strongest effect; high concentration (10-5mol / L) inhibited EPCs of these functions. The effect of simvastatin on proliferation was increased with time, while the adhesion and in vitro angiogenesis ability was the strongest at 48h. CONCLUSION: Simvastatin significantly increased the number of advanced EPCs and improved their function at low concentrations, while high concentrations inhibited it.