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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.
To study the expression of human basic fibroblast growth factor (bFGF) in acute myocardial ischemia model of rats and its effect on myocardial angiogenesis.Methods Human bFGF plasmid pcDNA / b was transfected into human Umbilical vein endothelial cells (HUVECs) were detected by MTT assay.PecDNA / b was transfected into 293 cells of human renal cell line and stably selected by G418 and detected by Western blotting.Acute myocardial infarction model was established in rats and normal saline and empty plasmid pcDNA3 respectively. 1 (+) and pcDNA / b were injected into infarct junction myocardium for 4 weeks and then harvested for routine HE staining and Masson staining to measure the number of microvessels and infarct size in each group.According to immunohistochemical staining and electron microscopy, Exogenous bFGF overexpression promoted the growth of HUVECs (pcDNA / b), which could efficiently express exogenous bFGF gene in 293 cells.A large number of neovascularization was observed at infarct junction of pcDNA / b injection group, and the total number of capillaries was significantly larger than that of control group The area was significantly smaller than the control group.pcDNA / b group in the infarct junction area bFGF positive.Electron microscopy showed that in the infarct junction of myocardial cells have capillaries proliferation, differentiated into two blood vessels.This experiment shows that, Overexpression of bFGF promotes capillary angiogenesis in acute ischemic myocardium of rats and provides an experimental basis for the study of bFGF gene therapy for ischemic cardiomyopathy.