人生长激素基因修饰的鼠骨骼肌成肌细胞移植对心肌梗死大鼠血流动力学的影响  

Hemodynamic effects of myoblast-based hGH gene therapy on myocardial infarction

作　　者：荣书玲[1,2] 王庸晋[2] 王晓林[2] 常超[1] 王裕勤[1] 高焱章[1] 米少华[1] 曹恒[1] 刘启云[1] 卢永昕[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院心内科,心血管病研究所,湖北武汉430022 [2]山西长治医学院附属和平医院,山西长治046000

出　　处：《中国病理生理杂志》2009年第1期42-47,共6页Chinese Journal of Pathophysiology

基　　金：国家高技术研究发展计划资助项目(“863”,No.2002AA745070);山西高校科技研究开发资助项目(No.200811034)

摘　　要：目的:探讨人生长激素(hGH)基因修饰的鼠骨骼肌成肌细胞移植对心肌梗死大鼠血流动力学和血管新生的影响。方法:结扎大鼠左冠状动脉前降支制作大鼠心力衰竭模型。2周后将冠脉结扎后存活的30只大鼠随机分为3组,hGH基因修饰的成肌细胞移植组(hGH组)、绿色荧光蛋白(GFP)基因修饰的成肌细胞移植组(GFP组)、注射等体积培养液的对照组。治疗4周后,血流动力学检查各组心功能指标;心肌组织进行HE染色检测心肌梗死面积,Ⅷ因子免疫组织化学检测血管新生。RT-PCR检测bax和bcl-2 mRNA的表达。Western blot-ting检测各组心肌hGH、血管内皮生长因子(VEGF)、caspase-3蛋白表达情况。结果:(1)与对照组和GFP组相比:hGH组血流动力学指标明显改善,心肌梗死面积缩小。(2)心肌组织Ⅷ因子免疫组织化学检查结果显示:hGH组血管密度显著高于GFP组和对照组。(3)RT-PCR检测结果显示hGH组bax mRNA水平显著低于GFP组和对照组,bcl-2 mRNA水平显著高于GFP组和对照组。(4)Western blotting检测结果显示hGH组大鼠心肌组织有hGH蛋白表达,其余两组心肌组织无hGH蛋白表达。与其它两组相比,hGH组caspase-3蛋白表达降低,VEGF蛋白表达增加。结论:hGH基因修饰的成肌细胞移植可以抑制细胞凋亡,与单独成肌细胞治疗相比可以诱导更大的血管化,更好地缩小心肌梗死面积,并改善心肌梗死大鼠血流动力学。成肌细胞治疗联合hGH基因治疗为心肌梗死后心力衰竭的治疗提供了一个新的途径。AIM： It has been showed that growth hormone （GH） exerts a favorable effect on cardiovascular function in clinical and animal studies. Skeletal myoblast transplantation has been shown to improve cardiac function in heart failure models by regenerating muscle. This study was to explore the effects of transplanted myoblasts transfected with human GH gene on hemodynamic indexes and angiogenesis of myocardial infarction in rats. METHODS ： Primary rat skeletal myoblasts were transfected with plasmids encoding human growth hormone （hGH） and green fluorescence protein （GFP）. Syngeneic adult rats underwent ligation of the left anterior descending coronary artery to establish MI models. Two weeks later, 6 × 10^6 transfected myoblasts with hGH gene, transfected myoblasts with GFP gene, or culture medium were transplanted into the infarcted myocardium as hGH group, GFP group or control group, respectively. Four weeks after injection, the evaluation of cardiac function in the survival rats was determined by physiological recorder. Histology was observed based on quantification of heart infracted size, myocardial fibrosis, and capillary density. RT - PCR was used to examine the mRNA expression of bax and bcl - 2. The protein expressions of hGH, vascular endothelial growth factor （VEGF） and caspase- 3 in myocardium were examined by Western. blotting. RESULTS： （1） Primary rat skeletal myoblasts were successfully transfected with hGH and GFP gene. The expression of hGH in transfected myoblast was demonstrated with immunohistochemistry and radioimmunoassay. （2） The-laemodynamics was improved significantly in hGH group and GFP group after four weeks therapy. Heart infracted size in hGH group was decreased significantly compared to GFP group and control group after therapy. （3） Capillary density in hGH group was significantly greater than that in both GFP group and control group （P 〈 0. 05）. （4） Reverse transcription polymerase chain reaction （ RT - PCR） showed that in hGH group mRNA

关 键 词：心力衰竭 充血性 成肌细胞 生长激素 基因疗法 

分 类 号：R363[医药卫生—病理学]