辛伐他汀影响内皮祖细胞数量黏附及凋亡的研究.doc

辛伐他汀影响内皮祖细胞数量、黏附及凋亡的研究 　　作者：周亚林,杨乃全,方翔，张雅莉，朱源生，梅介平 作者单位：徐州医学院附属淮安第二人民医院心脏内科，江苏淮安 　　【摘要】 目的 细胞水平观察不同浓度辛伐他汀对内皮祖细胞(endothelial progenitor cells，EPCs)功能的影响。方法 采用密度梯度离心法从人外周血获取单个核细胞，将其接种在人纤维连接蛋白包被培养板上，培养7天后，收集贴壁细胞，通过激光共聚焦显微镜鉴定，FITC-UEA-I和DiI-acLDL双染色阳性细胞为正在分化EPCs。以不同浓度辛伐他汀(分别为0.0001、0.001、0.01、0.1、1.0 Μmol/L)和EPCs培养。分别采用黏附能力测定实验、碘化丙啶(PI)标记观察辛伐他汀对EPCs黏附与凋亡的影响。结果 辛伐他汀显著提高了外周血EPCs的数量、黏附能力，并能抑制其凋亡。辛伐他汀浓度在0.01 Μmol/L时对EPCs的数量、黏附功能和抗凋亡能力影响达到最大。随着药物浓度的继续增大，EPCs的上述功能反呈下降趋势，但0.1 Μmol/L组仍高于对照组。结论 辛伐他汀能增加EPCs数量、黏附能力并能抑制其凋亡。 　　【关键词】 他汀类药物;内皮祖细胞;黏附;凋亡 　　Study on the effects of simvastatin on the number, adhesion and apoptosis 　　of endothelial progenitor cells 　　ZHOU Yalin,YANG Naiquan, FANG Xiang, ZHANG Yali, ZHU Yuansheng, MEI Jieping 　　(Department of Cardiovascular Diseases, The Affiliated Huaian Second Hospital of Xuzhou Medical College, 　　Huai′an, Jiangsu 223001, China) 　　Abstract： Objective To investigate the effects of simvastatin on EPC number, adhesion and apoptosis. Methods Total mononuclear cells (MNCs) isolated from human peripheral blood by Ficoll density gradient centrifugation were plated on fibronectin-coated culture dishes. After 7 days of culture, attached cells were obtained and assessed with a laser scanning confocal microscope, and differentiating EPCs were characterized as adherent positive cells by double staining of DiLDL-uptake and lectin binding. EPCs were treated with simvastatin of different concentrations (0.0001, 0.001, 0.01, 0.1 and 1.0 Μmol/L). EPC adhesion assay was performed by adherent cell counting on fibronectin-coated culture dishes. EPC apoptosis was evaluated by flow cytometry. Results Simvastatin promoted EPC number and adhesive capacity and prevented apoptosis . Simvastatin at 0.01 Μmol/L had the maximum effects on EPCs number, adhesion and anti-apoptosis. While the concentration of simvastatin further increased more than 0.01Μmol/L, the above improvement effects showed a tendency to decline, but the effects at