肌肉转录调节因子和连接蛋白43转基因成纤维细胞移植对心肌梗死大鼠脑钠素及梗死面积的影响  

Effect of myocardial transplantation after myogenic determination and connexin 43 genes transfected into dermal fibroblasts on B-type natriuretic peptide and infarcted size in rats with myocardial infarction

作　　者：汪进益[1] 范慧敏[1] 卢蓉[1] 张旭敏[1] 马亮[2] 李杨[2] 刘中民[1] 

机构地区：[1]同济大学附属东方医院心胸外科,上海市200120 [2]同济大学附属东方医院细胞治疗室,上海市200120

出　　处：《中国组织工程研究与临床康复》2008年第38期7499-7502,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：国家自然科学基金资助项目(30471719);上海市教委曙光计划(04SG25);上海市东方医院科研项目(DF2007Q07)~~

摘　　要：背景:研究表明基因工程将成纤维细胞改造为可兴奋细胞是可能的;而真皮成纤维细胞是机体内数量最大的"种子细胞库"。目的:观察肌肉转录调节因子(myogenic determination,MyoD)和连接蛋白43(connexin43,Cx43)基因共转染真皮成纤维细胞后在心肌梗死部位原位移植对急性心肌梗死大鼠血浆脑钠素浓度和心肌梗死面积的影响。设计、时间及地点:随机对照动物实验,于2006-06/2007-03在同济大学附属东方医院细胞治疗室和上海中医药大学动物实验中心完成。材料:雄性SD大鼠60只随机分成正常对照组、心肌梗死对照组、正常移植组和心肌梗死移植组,每组15只。方法:结扎左冠状动脉前降支制备心肌梗死模型。4周后再次开胸在自结扎远端紧靠结扎处,心肌梗死区与非梗死区交界处用微量注射器将100μL经BrdU标记的MyoD/Cx43-FB细胞悬液(细胞数1.0×106)注入左冠状动脉前降支,在结扎远端已缺血的心肌上用同样的针头于靠近心尖部、室间隔部、左室前侧壁缺血的心肌上多点多位置将另外100μL移植细胞悬液注入到心肌内,深度约4mm;对照组同法等量注入未含细胞的DMEM细胞培养液。主要观察指标:细胞移植前和移植后1d,1,2,4周动态检测血浆脑钠素浓度;同时监测各组大鼠心率、动脉压、左心室内压及左心室等容收缩期室内压最大上升速率和下降速率等血流动力学指标的变化;4周后处死大鼠取心肌标本染色后测定心肌梗死面积。结果:①免疫组织化学检测结果显示心肌瘢痕区边缘有BrdU阳性细胞存在。②细胞移植可显著改善心肌梗死大鼠的血流动力学。③心肌梗死对照组和心肌梗死移植组大鼠的血浆脑钠素浓度较正常对照组和正常移植组明显升高(P<0.01);在移植后1d、1周、2周心肌梗死移植组与心肌梗死对照组血浆脑钠素浓度差异无显著性意义,4周后明显降低(P<0.01)。④心肌梗死移植组�BACKGROUND： Recent study has proved the feasibility of transforming fibroblasts to excitable cells by means of gene engineering; dermal fibroblasts is the most abundant seed cells in organism. OBJECTIVE： To investigate the effects of dermal fibroblasts transfected by both myogenic determination and connexin 43 genes on B-type natriuretic peptide （BNP） and myocardial infarction （MI） size by its function of transplantation in situ at the lesioned sites in acute MI rats. DESIGN, TIME AND SETTING： Randomized control experiments were carried out in the Cell Treatment Centre, East Hospital Affiliated to Tongji University and Experimental Animal Center of Shanghai University of Traditional Chinese Medicine （Shanghai, China） from June 2006 to March 2007. MATERIALS： Sixty male Sprague-Dawley rats were randomly divided into four groups （n= 15）： normal control group, MI control group, normal transplantation group and MI transplantation group. METHODS： MI model was established by ligating left coronary artery anterior descending branch in rats. Four weeks later, thoracotomy was performed to conduct myocardial cell transplantation in situ. Dermal fibroblasts were converted by both myogenic determination and connexin 43 genes, then were labeled with bromodeoxyuridine. The prepared cell suspension, 100 mL containing 1.0×10^6 cells, was transplanted into the juncture between MI area and unlesioned area. Ischemic myocardium from surrounding cardiac apex, interventricular septum and left ventricular anterior lateral wall was also transplanted with 100 μL cell suspension, with the injector depth of 4 mm. In the control groups, DMEM cell liquid without cells were given. MAIN OUTCOME MEASURES： Before cell transplantation and 1 day, 1 week, 2 weeks, 4 weeks after cell transplantation, plasma BNP concentrations were detected. Heart rate, arterial blood pressure, left ventricular pressure, isovolumic contractive and diastolic function （left ventricular pressure maximal rate of rise and fall） were dete

关 键 词：MyoD基因 CX43基因 成纤维细胞 心肌梗死 脑钠素 心肌梗死面积 

分 类 号：R394.2[医药卫生—医学遗传学]