通心络体外促进人外周血内皮祖细胞的增殖、迁移、黏附的研究  

Experimental study on the effect of Tong Xin Luo on the proliferation,migration and adhesion of human endothelial progenitor cells from peripheral blood

作　　者：梁小卫[1] 田福利[1] 梁春[2] 吴宗贵[2] 

机构地区：[1]解放军二五一医院心内一科,张家口075000 [2]第二军医大学附属长征医院心内科,上海200003

出　　处：《国际心血管病杂志》2009年第3期184-187,190,共5页International Journal of Cardiovascular Disease

基　　金：国家重点基础研究发展计划(973计划)(2005CB523309)

摘　　要：目的:研究中药通心络对体外培养的人外周血内皮祖细胞(EPCs)增殖、迁移、黏附的影响。方法:采用密度梯度离心法分离培养人外周血单个核细胞,经FITC-UEA-I和Dil-acLDL双染色鉴定为正在分化的EPCs。进一步采用流式细胞仪检测其表面标志CD34、CD133,采用MTT比色法、transwell小室、细胞计数法分别观察50、100、200、500、750和1000μg/ml通心络超微粉溶液和500μg/ml通心络超微粉溶液作用0、6、12、24和36h后,对EPCs增殖、迁移、黏附的影响。结果:不同水平的通心络均改善了EPCs的增殖、迁移、黏附的功能,且在500μg/ml时作用最为显著。采用500μg/ml的通心络进行时效作用的研究显示,通心络呈时间依赖性地增强EPCs增殖、迁移、黏附能力,在36h达到高峰(P<0.01)。结论:通心络能显著改善外周血内皮祖细胞增殖、迁移、黏附能力,这可能是通心络防治心脑血管疾病的新机制。Objective： To explore the effects of Tong Xin Luo （TXL） on the proliferation.mi gration and adhesion of endothelial progenitor cells （EPCs） from human circulating blood in vitro. Methods：The TXL solution was prepared through ultrasonication according to the pervious literature. The rnononuelear cells （MNCs） were isolated from peripheral blood with Ficoll by density gradient centrifugation. MNCs were suspended in Medium 199 with 20% fetal blood serum （FBS） and vascular endothelial growth factor （VEGF）. After being cultured for 7d, the attached cells were characterized as Di-LDL uptaking and FITC-tectin binding by laser confocal microscope, and further demonstrated By expressing CD34 and CD133 using flow cytornetry. Cultured EPCs were incubated with TXL in a series of concentrations （0, 50, 100, 200, 500, 750, and 1000 μg/ml） for different durations （0, 6, 12, 24 and 36 hours）. The ceil morphology was observed and cell proliferation was determined by MTT assay, cell migration ability was examined by transwell chamber assay, the adhesion activity was determined by cell counts. Results： Incubation of EPCs with TXL increased the proliferative migration and adhesion ability of EPCs at different concentration, and this effect was most prominent when TXL concentration was 500μg/ml for 36 hours （P〈0.01）. Conclusion： the proliferation, migration and adhesion of EPCs were significantly improved by TXL.

关 键 词：通心络 内皮祖细胞 增殖 迁移 黏附 

分 类 号：R285[医药卫生—中药学]