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高表达ANG基因的自体骨髓基质细胞移植治疗猪缺血性心脏病
徐驯宇,陈前顺,黄郴,林兴,黄盛东,韩涛
0
(福建省立医院胸外科 福建医科大学省立临床医学院;第二军医大学附属长海医院心胸外科 上海 200433)
摘要:
目的 在猪的慢性心肌缺血的动物模型中移植其自体高表达ANG基因的骨髓基质细胞,分别观察其血管新生的效果及对心功能的改善作用。方法 建立猪的慢性缺血性心脏病动物模型;随机分3组:转染血管生长素基因的自体骨髓基质细胞移植组(组I);单纯自体骨髓基质细胞移植组(组II);注射无血清DMEM培养基的对照组(组III)。体外培养的自体骨髓基质细胞,利用Ad.ANG转染培养细胞,标记细胞;移植于模型猪心脏的缺血区域。观察指标包括:冠脉造影Rentrop分数、心脏射血分数、梗塞区的百分比、荧光显微镜下标记细胞、血管数量及ANG的表达和分泌。 结果 转染血管生长素基因的自体骨髓基质细胞移植组的冠脉造影Rentrop分数,心脏射血分数显著性增高;缺血心肌血管计数增高,可见大量CM-DiI标记的移植成活细胞,梗塞区面积缩小,ANG蛋白表达增高。结论 高表达ANG基因的自体基质细胞移植慢性缺血心肌中,移植细胞成活率高,局部血运和心功能改善明显。
关键词:  缺血性心脏病  自体骨髓基质细胞  血管生长素  细胞移植  腺病毒
DOI:10.3724/SP.J.1008.2014.00586
投稿时间:2013-12-06修订日期:2014-05-11
基金项目:福建省医学创新基金(2012-CX-5)
Effect of autologous angiogenin-overexpressing marrow stromal cells transplanted in a porcine model of chronic ischemia
Xu Xun-yu,CHEN Qian-shun,Huang Chen,LIN Xing,Huang Shengdong,HAN Tao
(Department of Thoracic Surgery, Fujian Provincial Hospital; Provincial Clinical Medical College of Fujian Medical University, Fuzhou 350001,,Fujian Province,China;;Department of Cardiothoracic Surgery, Shanghai Changhai Hospital, Second Military Medical University, Shanghai 200433 China)
Abstract:
Objective: To investigate the effect of autologous angiogenin-overexpressing marrow stromal cells transplanted in a porcine model of chronic ischemia. Methods: Standard experimental pigs underwent placement of a left circumflex artery ameroid occluder and were randomized to treatment with marrow stromal cells transfected ex vivo by angiogenin gene (Group I, n=11), or with marrow stromal cells without any gene transfected (Group II, n=10), and an additional group of animals served as a control (Group III, n=10). Cells were cultured, ex vivo angiogenin gene transferred, labeled by CM-DiI and transplanted by directly operating epicardial injection . Rentrop score were evaluated by angiography analysis, and ejection factor by echocardiography after 4 weeks of the therapy. The percentage of infarct area, vessel density, angiogenin protein expression, CM-DiI labeled cells were also evaluated at the same time. Results: Group I revealed significantly greater improvement in the Rentrop score, ejection factor and showed the smaller infarction area and apparent more normal gross left ventricular geometry and wall thickness than the other groups. Di-I labeled cells were observed on fluorescence microscopy indicated the successful engraftment of marrow stromal cells in group I in contrast to the Group II, which showed only sporadic labeled cells. In Group I the protein expression of angiogenin by Western blot and vessel number were also significantly increased. Conclusion: Transplantation of autologous angiogenin-overexpressing marrow stromal cells increases the survival of implanted cells and improves the impair heart function.
Key words:  Ischemic heart disease  Marrow stromal cells  Angiogenin  Cell transplantation  Adenovirus