巴曲酶可影响人外周血内皮祖细胞定向分化及增殖能力

背景：近年来内皮祖细胞促进机体血管新生方面的重要作用已形成共识,但内皮祖细胞的定向分化和扩增的问题是影响其疗效和广泛开展的主要原因。目的：观察人外周血内皮祖细胞体外分离、纯化、扩增和诱导分化为内皮细胞的可行性。方法：用密度梯度离心法从外周血获取单个核细胞,接种于包被有人纤维连接蛋白的细胞培养板中,培养6d后,收集贴壁细胞。以血管内皮细胞生长因子和碱性成纤维细胞生长因子对贴壁细胞进行诱导培养。另外,分别以0.05,0.1,0.2BU/mL巴曲酶培养贴壁细胞,观察内皮祖细胞的增殖能力。结果与结论：经血管内皮细胞生长因子和碱性成纤维细胞生长因子诱导后的细胞形态上表现内皮细胞特征,经流式细胞仪测定表达血管内皮细胞特有表面标志CD31和vWF,说明诱导后的细胞为内皮细胞。巴曲酶在体外培养条件下可增加血内皮祖细胞的数量,以0.1BU/mL浓度作用显著,且作用时间与血内皮祖细胞增殖能力的改善呈正相关。

Batroxobin influences the orientated differentiation and proliferation of endothelial progenitor cells from human peripheral blood in vitro

BACKGROUND：In recent years,it has been known that endothelial progenitor cells（EPCs） promote the angiogenesis of the body,but the directed differentiation and proliferation of EPCs limits their efficacy and widespread application.OBJECTIVE：To investigate the feasibility of inducing human peripheral blood derived EPCs into endothelial cells in vitro,and to test their phenotype and function.METHODS：Mononuclear cells（MNCs） were isolated from the peripheral blood by using Percoll density gradient centrifugation.The cells were then plated on fibronectin-coated culture dishes.After being cultured for 6 days,the attached cells were collected.EPCs were characterized as adherent cells double positive for DiI-LDL-uptake and lectin binding by direct flurescent staining under a laser scanning confocal microscope.The attached cells were cultured and induced by vascular endothelial growth factor（VEGF） and basic fibroblast growth factor（bFGF） in vitro.The morphological changes of the cells were observed.The cells were enumerated by flow cytometry（FCM） with CD31 monoclonal antibody and vWF monoclonal antibody.The attached cells were stimulated with DF-5201（final concentration：0.05,0.1,0.2 BU/mL） for the respective time points（6,12,24 and 48 hours）.Proliferation ability of EPCs was assayed by MTT assay.RESULTS AND CONCLUSION：After induced by VEGF and bFGF in vitro,EPCs became endothelial cell like cells and CD31 and vWF were expressed.Incubation of isolated human MNCs with DF-521 dose-and time-dependently increased the number of EPCs.EPCs can differentiate into endothelial cells by the induction of VEGF and bFGF.Incubation of EPCs with DF-521 increased the proliferative ability at different concentration.