辛伐他汀对骨髓源性内皮祖细胞动员及迁移的影响

目的研究辛伐他汀对兔内皮祖细胞（endothelial progenitor cells,EPCs）迁移、动员的影响。方法将2只兔从髂骨抽取骨髓,采用贴壁法,在条件培养基M199中培养EPCs,并用免疫荧光法鉴定CD34、CD133、VEGFR-2。经鉴定的EPCs,在不同浓度辛伐他汀（分别为0.01,0.1,1.0μmol/L）作用下,Transwell实验观察辛伐他汀对EPCs迁移的影响。6只兔随机分为实验组和对照组,每组3只。6只兔颅骨造极限骨缺损模型,实验组和对照组在颅骨缺损处分别植入辛伐他汀复合聚乳酸薄饼或聚乳酸薄饼,术后10天,取外周血,流式细胞仪检测CD34/CD133双阳性细胞表达率,观察辛伐他汀对EPCs动员的效果。结果Transwell实验,辛伐他汀作用于EPCs16小时,0.1μmol/L和1.0μmol/L辛伐他汀组细胞迁移数量OD值（0.097±0.011,0.099±0.019）较0.01μmol/L辛伐他汀组（0.075±0.013）与对照组（0.077±0.014）多,差异有显著性（P〈0.05）。造模术后10天,辛伐他汀组3只兔外周血中CD34^＋/CD133^＋细胞表达率为0.28%,0.84%,0.28%,对照组为0.26%,0.11%,0.09%。结论辛伐他汀可动员EPCs到外周血,可提高骨髓源性EPCs的迁移能力。

Effect of Simvastatin on Mobilization and Migration of Endothelial Progenitor Cells

Objective To investigate the effect of simvastatin on the mobilization and migration of endothelial progenitor cells（EPCs）.Methods EPCs were harvested from the bone marrows of two rabbits,cultured with M199,and identified by immunohistochemistry.The identified EPCs were then treated with simvastatin with different concentrations（0,0.01,0.1,1.0 μmol/L）,and their migration induced by simvastatin was determined with Transwell chamber assay.Six rabbits models of cranial bone defect were established and divided into control and experiment groups（3 in each）.In order to elicit the effects of simvastatin on mobilization of EPCs,simvastatin was embedded in polylactic acid compound,and implanted into the cranial bone defect area in the experiment group.Meanwhile,polylactic acid was implanted in the control animals.After 10 days,the expression rate of CD34^＋/CD133^＋ EPCs in the rabbit peripheral blood was counted by flow cytometry to determine the motivating effect of simvastatin.Results In Transwell experiment,16 hours after adding simvastatin（0,0.01,0.1 or 1.0 μmol/L）,the cell migration ability was obviously increased showing a dose-dependent trend（OD value：0.077±0.014 in control group and 0.075±0.013 in 0.01 μmol/L group vs 0.097±0.011 in 0.1 μmol/L group and 0.099±0.019 in 1.0 μmol/L group,P0.05）.In animal experiment,10 days after the implantation of simvastatin,the expression rate of CD34^＋/CD133^＋ EPCs were 0.28%,0.84%,and 0.28% respectively in the three rabbits,while in the control groups,the rates were 0.26%,0.11% and 0.09%.Conclusion Simvastatin could mobilize EPCs into peripheral blood,and improve migration capability of EPCs.