血管内皮生长因子对大鼠骨髓内皮祖细胞生物学特性的影响

Effects of Vascular Endothelial Growth Factor on the Biological Characteristics of Rat Bone Marrow Endothelial Progenitor Cells

目的观察内皮祖细胞(EPC)在不同浓度血管内皮生长因子(VEGF)作用下的生物学特性,探讨VEGF对EPC生物学特性的影响。方法采用密度梯度离心法获取鼠骨髓单个核细胞,培养7d后收集贴壁细胞鉴定EPC,用含不同浓度VEGF(0、10、50ng/ml)的培养基继续培养。采用MTT比色法、Transwell小室实验观察EPC的增殖和迁移能力,应用纤维蛋白凝胶观察EPC体外血管形成能力,流式细胞术检测EPC的分化能力。结果培养7d的细胞CD133和CD34阳性率分别为69.44%和81.05%。MTT实验显示,10ng/ml组细胞的吸光值明显大于0ng/ml组和50ng/ml组,0ng/ml组的吸光值明显大于50ng/ml组(P<0.05);迁移实验、体外血管形成实验及诱导分化实验显示,50ng/ml组和10ng/ml组较0ng/ml组更能促进EPC的迁移、体外血管形成及诱导分化,且50ng/ml组强于10ng/ml组(P<0.05)。结论 VEGF能够提高EPC迁移能力、体外血管形成能力及促进诱导分化,且随着VEGF浓度的升高而增强。VEGF能够提高EPC增殖能力,但随着浓度的升高而减弱。

Objective To investigate the effect of vascular endothelial growth factor（VEGF）on the biological functions of rat bone marrow endothelial progenit cells （EPC）.Methods Total mononuclear cells （MNCs）were extracted from rat bone marrow by density gradient centrifugation.After cultured for 7 d,the attached cells were collected and incubated with VEGF in a series of concentrations （0 ng/ml,10 ng/ml,50 ng/ml）.The EPC＇s capability of proliferation,migration,in vitro vasculogenesis,and differentiation into endothelial cell were evaluated with MTT assay,Transwell chamber assay,Fibrin-gel and flow cytometry,respectively.Results At 7 days after culture,the positive rates of CD133 and CD34 were 69.44%and 81.05%;MTT assay results showed the OD value of 10 ng/ml group was higher than that of the 0 ng/ml and the 50 ng/ml groups significantly,while the 0 ng/ml group was higher than that of the 50 ng/ml group（P 〈 0.05）;migration,in vitro vasculogenesis and differentiation analysis results demonstrated that the capability of promoting the EPC migration,in vitro vascularization and the differentiation,compare in 50 ng/ml and 10 ng/ml groups was stronger than 0 ng/ml group,while the capability of 50 ng/ml group was stronger than 10 ng/ml group （P 〈 0.05）.Conclusion VEGF could promote the migration,in vitro vasculogenesis and differentiation of EPC dose-dependently.VEGF could also promote the proliferation of EPC,but the higher the concentration of VEGF was,the promoting effects the weaker.