过表达bFGF促进大鼠急性缺血心肌的毛细血管生成  

Overexpression of bFGF Promotes Angiogenesis in Rats with Acute Myocardial Ischemia

作　　者：王雅梅[1] 刘冰[2] 孙丽翠[1] 祁雅慧[2] 司杨[1] 闫豫东[2] 郑少鹏[1] 

机构地区：[1]首都医科大学生物化学与分子生物学系,北京100069 [2]首都医科大学医学实验测试中心,北京100069

出　　处：《中国生物化学与分子生物学报》2011年第1期78-83,共6页Chinese Journal of Biochemistry and Molecular Biology

基　　金：北京市科委科技合同项目(No.954024800);北京市教育委员会资助项目(No.KM200310025094)~~

摘　　要：研究过表达人碱性成纤维细胞生长因子(basic fibroblast growth factor,bFGF)在大鼠急性心肌缺血模型中的表达及其对心肌血管再生的影响.将含人bFGF质粒pcDNA/b转染人脐带静脉内皮细胞(HUVEC),进行MTT检测.pcDNA/b转染人肾细胞293细胞系,进行G418稳定筛选,Western印迹检测.建立大鼠急性心肌梗死模型,分别将生理盐水、空质粒pcDNA3.1(+)和pcDNA/b分3点注射于梗死交界处心肌内4周后取材,做常规HE染色和Masson染色,测量各组微血管数量和梗死面积.经免疫组织化学染色鉴定和电镜观察,过表达外源bFGF可以促进HUVEC(细胞)生长;pcDNA/b能在293细胞中高效表达外源bFGF基因.pcDNA/b注射组梗死交界处可见大量新生血管,毛细血管总数明显大于对照组,梗死面积明显小于对照组.pcDNA/b组在梗死交界区有bFGF阳性表达.电镜观察显示,在梗死交界处心肌细胞间有毛细血管增生,分化成2个血管腔.本实验证明,过表达bFGF具有促进大鼠急性缺血心肌的毛细血管生成的作用,为bFGF基因治疗缺血性心肌病的研究提供实验基础.The overexpression of recombinant human basic fibroblast growth factor（bFGF） in the rats with acute myocardial ischemia and its effect were detected on myocardial angiogenesis.The recombinant plasmid pcDNA /b containing human bFGF cDNA was transferred into HUVEC mediated by liposome,then detected by MTT.pcDNA/b was transfected into 293 cells mediated by liposome.The positive cellswere selected and determined by Western blot.Established model rats with acute myocardial ischemiawere divided randomly into three groups,and saline,50μL naked plasmid DNA or pcDNA/b wereintramyocardially injected at three sites into the border zone of infarction.The hearts of rats were excisedto evaluate the density of the capillary vessels and infarct size,HE staining,Masson＇s trichrome staining,immunohistochemical staining and electron microscopy after were done at four weeks.The results showedthat overexpressed bFGF protein has strong biological activity to stimulate HUVEC proliferation.ThebFGF protein was expressed successfully in 293 cells.In pcDNA/b group,myocardial cells in infarctionborder zone showed bFGF expression in cytoplasm,the other groups showed negative.The density ofmyocardial capillary vessels in pcDNA/b group was significantly more than those in control groups.Therewas more visible hyperplasia of vascular endothelium in pcDNA/b group but less in the other two groups.In this study,human bFGF was cloned and its protein was expressed successfully in vitro.Overexpressionof bFGF promotes angiogenesis in model rats with acute myocardial ischemia.It provides a basis for thefurther study of biological functions of human bFGF.

关 键 词：碱性成纤维细胞生长因子 真核表达载体 基因过表达 心肌梗死 

分 类 号：Q785[生物学—分子生物学]