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本文旨在探讨他汀类药物氟伐他汀对外周血内皮祖细胞(endothelial progenitor cells,EPCs)微量和功能的影响。用密度梯度离心从外周血获取单个核细胞,将其接种在人纤维连接蛋白(human fibronectin)包被的培养板中,培养7d后,收集贴壁细胞,加入不同浓度氟伐他汀(分别为0.01、0.1、1、10μmol/L)和辛伐他汀(1μmol/L),培养一定的时间(6、12、24、48h)。用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的EPCs,用流式细胞仪检测其表面标志进一步鉴定EPCs,在倒置荧光显微镜下计数。采用MTT比色法、改良的Boyden小室、粘附能力测定实验和体外血管生成试剂盒观察EPCs的增殖能力、迁移能力、粘附能力和体外血管生成能力。结果显示,氟伐他汀可显著增加外周血EPCs的数量,并且EPCs数量随氟伐他汀浓度增加及作用时间延长而增加,1μmol/L浓度氟伐他汀作用24h对EPCs的数量影响最为显著(较对照组增加1.5倍,P<0.05)。在动物实验中,喂养氟伐他汀3周后,大鼠的EPCs也较对照组增加2倍(P<0.05),进一步支持了体外实验的结果。氟伐他汀和辛伐他汀也显著改善外周血EPCs的粘附能力、迁移能力、增殖能力和体外血管生成的能力,相同浓度的氟伐他汀和辛伐他汀(1μmol/L)对EPCs数量和功能的影响并无显著差异。上
This article aims to investigate the effect of statin fluvastatin on the trace and function of peripheral blood progenitor cells (EPCs). Mononuclear cells were obtained from peripheral blood by density gradient centrifugation and seeded on human fibronectin-coated plates. After 7 days of culture, adherent cells were collected and different concentrations of fluvastatin (0.01 , 0.1,1,10μmol / L) and simvastatin (1μmol / L) for a certain period of time (6,12,24,48h). Confocal laser scanning microscopy was used to identify the differentiated EPCs positive for FITC-UEA-I and DiI-acLDL double stained cells. The surface markers of FITC-UEA-I and DiI-acLDL were identified by flow cytometry to further identify EPCs and count under inverted fluorescence microscope. MTT assay, modified Boyden chamber, adhesion assay and in vitro angiogenesis kit were used to observe the proliferation, migration, adhesion and in vitro angiogenesis of EPCs. The results showed that fluvastatin can significantly increase the number of EPCs in peripheral blood, and the number of EPCs increased with the increase of fluvastatin concentration and prolonged action time. Fluvastatin at a concentration of 1μmol / L for 24h had the most significant effect on EPCs Group increased 1.5 times, P <0.05). In animal experiments, 3 weeks after feeding fluvastatin, EPCs in rats also increased by 2-fold (P <0.05) compared with the control group, further supporting the results of in vitro experiments. Fluvastatin and simvastatin also significantly improved EPCs adhesion, migration, proliferation and angiogenesis in vitro. Fluvastatin and simvastatin (1μmol / L) at the same concentration significantly reduced the number and function of EPCs There was no significant difference in the effects. on