灯盏花素对内皮祖细胞数量、功能及iNOS的影响  

Effects of Breviscapine on Number,Activity and iNOS of Endothelial Progenitor Cells

作　　者：陈图刚[1] 李鸣媛[1] 谭维羚[1] 马战清[1] 钟志英[1] 孙国锋[1] 周宇昕[1] 

机构地区：[1]南昌大学第四附属医院,江西南昌330003

出　　处：《实用中西医结合临床》2010年第4期1-3,共3页Practical Clinical Journal of Integrated Traditional Chinese and Western Medicine

基　　金：江西省科学技术厅课题(编号:2007A122)

摘　　要：目的:观察灯盏花素对外周血内皮祖细胞(endothelial progenitor cells,EPCs)数量、功能及诱导型一氧化氮合酶(iNOS)的影响。方法:采用密度梯度离心法从外周血获得单个核细胞,将其接种在人纤维连接蛋白包被的培养板上,培养7d后收集贴壁细胞,加入不同浓度灯盏花素(分别为5、10、20、40mg/L)培养24h,同时,对照组与灯盏花素20mg/L组分别培养6、12、24、48h。激光共聚焦显微镜鉴定FITC-UEA-I和Dil-acLDL双染色阳性细胞为正在分化的EPCs,将其在倒置荧光显微镜下计数。然后分别采用MTT比色法、改良的Boyden小室、黏附能力测定实验和体外血管生成试剂盒来观察EPCs的增殖能力、迁移能力、黏附能力和体外血管形成能力,免疫印迹杂交法半定量测定iNOS含量。结果:灯盏花素能增加外周血EPCs数量、增殖能力、迁移能力、黏附能力和体外血管形成能力,并且其数量、增殖能力、迁移能力、黏附能力和体外血管形成能力随灯盏花素浓度和作用时间的增加而增加,灯盏花素还能增加EPCs中iNOS含量。结论:灯盏花素增加EPCs的数量、增殖能力、迁移能力、黏附能力和体外血管形成能力,与其增加iNOS含量有关。Objective：To investigate whether Breviscapine has influences on number,activity,and inducible nitric oxide synthase（iNOS） of endothelial progenitor cells（EPCs）.Methods：Total mononuclear cells（MNCs） were isolated from peripheral blood by Ficoll density gradient centrifugation,and then the cells were plated on fibronectin-coated culture dishs.After 7 days cultured,attached cells were stimulated with Breviscapine（to make a series of final concentrations：5,10,20,40mg/L）for 24 hours,the control group and group Breviscapine 20mg/L cultured for 6,12,24,48h.EPCs were characterized as adherent cells double positive for DiLDL-uptake and lectin binding by direct fluorescent staining.EPCs proliferation,migration were assayed with MTT assay and modified Boyden chamber assay,respectively.EPCs adhesion assay was performed by replating those on fibronectin-coated dishes,and then adherent cells were counted.In vitro vasculogenesis activity was assayed by in vitro vasculogenesis kit.iNOS was assayed by western blot.Results：Incubation of isolated human MNCs with Breviscapine increased the number of EPCs,their proliferative,migratory,adhesive and in vitro vasculogenesis capacity in a concentration and time dependent manner.Breviscapine also increased EPCs＇iNOS.Conclusion：Breviscapine increases the number,proliferative,migratory,adhesive and in vitro vasculogenesis capacity of EPCs,the effects were related to the increment of iNOS by Breviscapine.

关 键 词：灯盏花素 内皮祖细胞 诱导型一氧化氮合酶 

分 类 号：R541.4[医药卫生—心血管疾病]