高同型半胱氨酸血症患者循环内皮祖细胞的变化  

Changes in the number and activity of endothelial progenitor cells from peripheral blood in patients with hyperhomocysteinemia

作　　者：朱军慧[1] 马彩艳[2] 陈君柱[3] 王兴祥[3] 张芙荣[3] 孙坚[3] 

机构地区：[1]浙江大学医学院附属邵逸医院心内科,杭州310016 [2]浙江省立同德医院心内科 [3]浙江大学医学院附属第一医院心内科

出　　处：《临床心血管病杂志》2008年第4期277-281,共5页Journal of Clinical Cardiology

基　　金：浙江省科学技术研究基金(No:2004C30012)

摘　　要：目的:观察高同型半胱氨酸血症(HHcy)患者外周血内皮祖细胞(EPCs)数量和功能的变化。方法:入选患者60例,其中HHcy患者30例(HHcy组),非HHcy患者30例(对照组)。采用全自动荧光偏振免疫分析法检测血浆同型半胱氨酸(homocysteine,Hcy),流式细胞仪计数循环EPCs(即AC133+KDR+细胞)。此外,采用密度梯度离心法从外周血获取单个核细胞,将其接种在人纤维连接蛋白包被培养板,培养4d后贴壁细胞进行细胞化学分析。激光共聚焦显微镜鉴定FITC标记荆豆凝集素Ⅰ和DiI标记的乙酰化LDL双染色阳性细胞为正在分化的EPCs。采用改良的Boyden小室和黏附能力测定实验观察EPCs的迁移和黏附能力。结果:HHcy患者循环EPCs数量、外周血正在分化的EPCs数量[(63.9±11.7)、(36.1±6.5)个]均较对照组的[(91.5±14.2)、(51.5±8.3)个]明显减少,均P<0.01;这2种EPCs数量均与血Hcy水平呈反向线性关系。HHcy患者EPCs的迁移和黏附能力也较对照组的明显受损,EPCs的这2种功能与血Hcy水平呈反向线性关系。结论:HHcy患者外周血EPCs数量减少、功能减退,且EPCs的数量和功能与血Hcy水平呈反向线性关系。Objective：To investigate changes in the number and activity of endothelial progenitor cells （EPCs） from peripheral blood in patients with hyperhomocysteinemia （HHcy）. Method： Human EPCs were isolated and cultured from patients with H Hcy （n=30） and matched volunteers （n= 30）. Circulating EPCs were enumerated as AC133^＋ KDR^＋ cells via fluorescence-activated cell sorter analysis. Additionally, EPCs were expanded from human blood in vitro and identified by Dil-acLDL uptake and lectin staining by direct fluorescent staining under a laser scanning confocal microscope. EPCs migration activities were determined by modified Boyden chamber assay. EPCs adhesion assay was performed by replating cells on fibronectin-coated dishes, then adherent cells were counted. Result：A significant decrease was observed in circulating EPCs （AC133^＋ KDR^＋ cells） numbers in patients with HHcy compared with control subjects （[63.9±11.7] vs [91.5±14.2] cells/ml blood, P〈0. 01）. And the numbers of EPCs also decreased in patients with HHcy after ex vivo cultivation （[36.1±6. 5] vs [51.5±8.3] EPCs/×200 field, P〈0. 01）. Both circulating EPCs and differentiated EPCs were inversely correlated with total Hcy levels. In addition, EPCs {rom patients with HHcy were significantly impaired in their migratory capacity and ability to adhere to fibronectin compared with controls. Conclusion： The present study demonstrated that EPCs numbers and functional capacity were impaired in patients with HHcy.

关 键 词：高同型半胱氨酸血症 内皮祖细胞 内皮 血管 动脉粥样硬化 

分 类 号：Q517[生物学—生物化学]