内脏脂肪素对内皮祖细胞血管形成能力的影响  

Effect of visfatin on angiogenic function of endothelial progenitor cells

作　　者：肖坚[1] 肖振军[2] 彭军[1] 

机构地区：[1]中南大学药学院药理学系,湖南长沙410078 [2]中南大学湘雅医院老年科,湖南长沙410008

出　　处：《中国现代医学杂志》2009年第10期1502-1505,1510,共5页China Journal of Modern Medicine

摘　　要：目的内脏脂肪素(visfatin)是一种能够促进血管新生的新型脂肪细胞因子。血管内皮生长因子(VEGF)是血管新生中非常关键的因子。本文在培养的内皮祖细胞(EPCs),探讨visfatin对EPCs血管新生的影响。方法密度梯度离心法分离人脐血的单个核细胞,利用EGM-2细胞培养基(含EPCs分化所需各种生长因子)进行培养,诱导单个核细胞贴壁向EPCs分化。流式细胞术测定早期EPCs干细胞分子标志CD133/CD34;Dil-ac-LDL和FITC-UEA-1双染鉴定正在分化的EPCs。不同浓度的重组visfatin(0.01-10nM)处理EPCs24h。Tube形成和迁移实验评价EPCs血管新生能力;荧光定量实时PCR检测VEGF mRNA水平。结果0.1nM和1nM的visfatin能增加EPCs形成管腔样结构(与对照组比,P<0.01),10nM的visfatin组EPCs形成管腔样结构能力和迁移能力却显著降低(与1nM visfatin组比,P<0.01)。0.1nM和1nM vis-fatin组细胞VEGF mRNA水平明显升高(与对照组比,P<0.01),而10nM visfatin组细胞VEGF mRNA明显降低(与1nM visfatin组比,P<0.01)。结论生理浓度visfatin能促进EPCs的血管形成,病理浓度其血管形成能力降低,其机制可能与VEGF的表达有关。[Objective] Visfatin, a new adipocytokine, has been reported to promote angiogenesis. Vascular endothelial growth factor （VEGF） expressed in endothelial progenitor eells, is thought as a novel modulator of angiogenesis. The aim of this study was to investigate the role of visfatin in angiogenie function in endothelial progenitor cells. [Methods] EPCs were isolated by density gradient centrifugation from human cord blood mononuelear ceils, and cultured in EBM-2 supplemented with EGM-2 Single-Quots. Adherent EPCs were characterized by dual staining for acetylated low-density lipoprotein （ac-LDL） and ulex europaeus agglutinin-1 （UEA-1）, and by flow cytometry for detecting the stem cell marker CD133 and CD34 or by immunofluorescent analysis. After incubation of EPCs with different concentrations of visfadn （0.01-10 nM） for 24 hours, we examined the capabilities of tube formation and cell migration in EPCs to judge the angiogenie effects of visfatin. Quantitative real time PCR was used to measure the expression of VEGF mRNA. [Results] visfatin at the concentrations of 0.1 nM and 1 nM could enhance cell tube formation and cell migration reflecting angiogenio capability of EPCs compared with control （P 〈0.01）, whereas the tube formation and cell migration was decreased at 10 nM compared with that atl nM （P 〈0.01）. Visfatin at 0.1 nM and 1 nM could upregulate the mRNA expression of VEGF compared with control （P 〈0.01）, whereas the expression of VEGF mRNA was decreased at 10 nM of visfatin compared with that at lnM （P 〈0.01）. [Conclusions] Visfatin at the physiological concentration could promote the angiogenic function of EPCs, whereas the anglogenic function of EPCs was a.ttenuated at the pathological concentration of visfatin. The effect of visfatin on EPCs an-giogenesis is related to regulating VEGF production.

关 键 词：内脏脂肪素 内皮祖细胞 血管内皮生长因子 血管新生 

分 类 号：R365[医药卫生—病理学]