目的 观察吡格列酮对外周血内皮祖细胞(EPCs)数量与增殖能力的影响.方法 采用Ficoll密度梯度离心法从外周血获得单个核细胞,将其接种于包被人纤维连接蛋白的培养板上,培养14d后收集贴壁细胞,加入不同浓度(10-6、10-5、10-4、10-3mmol/L)吡格列酮分别培养24、48、72h,同时采用激光共聚焦显微镜鉴定,FITC-UEA-I和Dil-acLDL双染色阳性细胞为正在分化的EPCs,并在倒置荧光显微镜下计数;后采用MTT比色法观察EPCs的增殖能力.结果 在作用48h后,不同浓度的吡格列酮均增加了外周血EPCs数量和增殖能力;10-5mmol/L吡格列酮作用48h时影响最为显著,EPCs数量和增殖能力(分别为54.0±2.9和0.593±0.033)明显高于对照组(分别为30.0±3.1和0.178±0.018),差异均有统计学意义(均P＜0.01).结论 吡格列酮可增加EPCs数量并改善EPCs的增殖能力.%Objective To investigate the effects of pioglitazone on the proliferation of peripheral endothelial progenitor cells (EPCs).Methods Total mononuclear cells (MNCs) were isolated from peripheral blood samples by Ficoll density gradient centrifugation, and then the cells were plated on fibronectin-coated culture dishes.After 14-day culture, adherent cells were treated with pioglitazone in a series of final concentrations of 10-6, 10-5, 10-4 and 10-3mmol/L for 24, 48 and 72h, EPCs were characterized as adherent cells double positive for DiLDL-uptake and lectin binding by direct fluorescent staining under a laser scanning confocal microscope.EPCs proliferation assayed by MTT assay,Results The number and the proliferation of EPCs were significantly increased by pioglitazone for 48 h and 72h as compared with that in control.Pioglitazone increased the number of EPCs( 54.0 ± 2.9vs 30.0 ± 3.1, P＜ 0.01 )and improved the proliferative capacity of EPCs( 0.593 ± 0.033 vs 0.178 ± 0.018,P＜ 0.01 )especially in concentration of 10-5mmol/L for 48 h.Conclusion The results suggest that pioglitazone may increase the number and the proliferative capacity of EPCs.
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