血管内皮生长因子调节内皮祖细胞生物学功能

Effects of vascular endothelial growth factor on biological functions of bone marrow-derived endothelial progenitor cells

摘要:

目的研究血管内皮生长因子(VEGF)对体外培养骨髓源性内皮祖细胞(EPCs)数量及增殖、迁移、黏附功能的影响及机制初探。方法密度梯度离心法获取骨髓单个核细胞,FITC-荆豆凝集素I、DiI-乙酰化低密度脂蛋白荧光双染鉴定。单个核细胞培养7d后分为对照组和VEGF干预组。VEGF干预组加入不同浓度VEGF(25,50,75,100μg/L)培养48 h,分别采用四氮唑溴盐比色法、改良的Boyden小室和黏附能力测定观察EPCs的增殖、迁移和黏附能力。RT-PCR法半定量检测VEGF对EPCs内皮型一氧化氮合酶(eNOS)mRNA表达的影响。硝酸还原酶法比色测定VEGF对EPCs分泌一氧化氮的影响。结果VEGF可浓度依赖性地增加EPCs数量并明显促进EPCs的黏附、迁移和增殖能力,与对照组比较差异显著。VEGF可上调EPCs eNOS mRNA的表达,促进EPCs分泌一氧化氮。结论VEGF可能通过上调EPCs eNOS mRNA的表达影响EPCs部分生物学功能。

Abstract:

Objective To observe the effect of vascular endothelial growth factor(VEGF) on endothelial progenitor cells(EPCs) and to elucidate its possible mechanism.Methods Mononuclear cells(MNCs) isolated from bone marrow by density gradient centrifugation combined with adherent cell filtration were plated on fibronectin-coated culture dishes.After culturing for 7d,adherent cells were cultured with different concentrations of VEGF(25,50,75,100 μg/L) for 48 h.EPCs proliferation and migration ability were observed and adhesion assay was performed.The endothelial nitric oxide synthase(eNOS) mRNA expression of EPCs was measured by RT-PCR.Results VEGF dose-dependently increased the number of EPCs and promoted EPCs proliferation,migration and adhesive capacity.VEGF could upregulate eNOS mRNA expression of EPCs.Conclusion VEGF might affect the biological functions of EPCs through upregulation of eNOS mRNA expression of EPCs.