辛伐他汀对内皮祖细胞增殖、迁移、凋亡及黏附影响的研究

【作者】 杨乃全；

【导师】 张馥敏；

【作者基本信息】 南京医科大学 ， 心血管内科， 2008， 硕士

【摘要】 目的:细胞水平观察不同浓度辛伐他汀对内皮祖细胞(endothelialprogenitor 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EPCs。以不同浓度辛伐他汀(分别为0.0001、0.001、0.01、0.1、1.0μM)或0.01μM辛伐他汀+PI3K阻断剂(LY294002)和EPCs培养24h。分别采用MTT比色法、改良的Boyden小室、流式细胞仪和黏附能力测定实验来观察辛伐他汀对EPCs的增殖、迁移、凋亡和黏附影响,并探讨LY294002干预作用。结果:辛伐他汀显著提高了外周血EPCs的增殖能力、迁移能力、抗凋亡能力和黏附能力(P＜0.05)。辛伐他汀浓度在0.01μM时对EPCs功能影响达到最大。随着药物浓度的继续增大,EPCs的上述功能反呈下降趋势,但0.1μM组仍高于对照组。辛伐他汀对EPCs的功能影响能被LY294002(PI3K抑制剂)阻断。结论:辛伐他汀能增加EPCs的增殖、迁移、抗凋亡和黏附能力,其机制可能与PI3K/Akt信号途径有关。更多还原

【Abstract】 Objective: To investigate in vitro whether simvastatins of different concentrations have influences on the functions of endothelial progenitor cells (EPCs).Methods: Total mononuclear cells (MNCs) were isolated from human peripheral blood by Ficoll density gradient centrifugation, and then the cells were plated on fibronectin-coated culture dishes. After 7 days cultured, attached cells were isolated and were assessed with a laser scanning confocal microscope, EPCs were characterized as cells double positive for DiLDL-uptake and lectin binding. EPCs were treated with simvastatins of different concentrations (0.0001、0.001、0.01、0.1、1.0μM),or LY294002(PI3K inhibitor)+ simvastatins(0.01μM) for 24h. The influences of Simvastatins on EPCs’ proliferation, migration , apoptosis were assayed with MTT assay, modified Boyden chamber assay and FACS assay, respectively. EPCs adhesion assay was performed by replating those on fibronectin-coated dishes, and then the adherent cells were counted.Results: Simvastatins strikingly improved the ability in proliferating, migrating, anti-apoptosis and adhering of EPCs isolated from human peripheral blood (p<0.05). Simvastatins at 0.01μM concentration reached the maximum effect on EPCs. While the concentration of simvastatins was further increased more than 0.01μM, the above improvement effects showed a tendency to decline, but the effects at 0.1μM concentration were still greater than those in control group. And the influence of Simvastatins on these functions of EPCs could be abolished by LY294002(PI3K inhibitor).Conclusion: The present study established that Simvastatins could improve EPCs’ proliferation, migration , anti-apoptosis and adhesion and the mechanism maybe involves the PI3K/AKt pathway.更多还原