移植表达δ-SG基因的骨髓间充质干细胞对TO-2型仓鼠心功能的影响  

Transplantation of Bone Marrow-derived Mesenchymal Stem Cells Expressing Hamsterδ-SG Gene for TO-2 Hamster Cardiac Repair

作　　者：黄海怡[1] 黄琳[1] 吴士尧[1] 陈元美[1] 

机构地区：[1]上海交通大学第九人民医院心血管内科,上海200011

出　　处：《中国分子心脏病学杂志》2011年第4期224-229,共6页Molecular Cardiology of China

基　　金：上海市科委基金资助项目(No.064119624)

摘　　要：目的采用基因与细胞相结合的新方法,研究将经重组腺相关病毒(rAAV)介导的δ-SG基因修饰的骨髓间充质干细胞(MSCs)植入缺失δ-SG基因的TO-2型仓鼠心肌内,对其心功能的影响。方法将遗传性DCM鼠(TO-2仓鼠)分为MSCs组(n=15)、rAAV-δ-SG组(n=15)、rAAV-δ-SG-MSCs组(n=15),心室壁注入移植细胞。移植后5w和10w,分别进行心超检测左室舒张末直径(LVDd)、左室收缩末直径(LVSd),舒张末室间隔厚度(IVSd),舒张末左室后壁厚度(LVPW),左室射血分数(LVEF);免疫组化测心肌细胞特异蛋白;RT-PCR检测心肌中δ-SG基因表达;免疫荧光检测细胞分化、迁移;电镜检测新生细胞。结果与rAAV-δ-SG、MSCs组相比较,rAAV-δ-SG-MSCs组LVDd、LVSd减小(P<0.05),EF、IVSd、LVPW增大(P<0.05),而rAAV-δ-SG与MSCs组相比较无统计学差异;rAAV-δ-SG-MSCs组心肌细胞较成熟,结构较完整,心肌细胞特异蛋白表达增多,毛细血管密度增加(P<0.01);电镜中,可见结构完整的心肌细胞及新生血管出现。结论 rAAV-δ-SG-MSCs能与宿主细胞有效结合,与rAAV-δ-SG、MSCs治疗相比较,能更有效地修复TO-2仓鼠心肌细胞肌膜骨架蛋白,持续修复受损心肌,改善心功能,延缓心腔扩大。Objective The aim was to investigate the effects of transplantation of bone marrow-derived mesenchymal stem cells expressingδ-SG gene into myocardium of δ-SG-gene-deleting TO-2 hamster. Methods: TO-2 hamsters were divided into 3 groups, MSCs group (n=15), rAAV-δ-SG group (n=15), and rAAV-δ-SG-MSCs group (n=15). Five and Ten weeks after cell transplantation into myocardium of TO-2 hamsters, heart function including the left ventricular end-diastolic diameter (LVDd), LV end-systolic diameter (LVSd), diastolic interventricular septal thickness(IVSd),diastolic left ventricular posterior wall thickness(LVPW)and ejection fraction (EF) were assessed by echocardiography. After hearts were harvested, δ-SG gene expression were detected by RT-PCR. And then, cardiac specific actin was detected by immunohistochemistry, and cell differentiation and migration were observed by immunofluorescence staining. Finally, newborn cells were observed by electron-microscope.Results: As compared with MSCs and rAAV-δ-SG groups, echocardiography showed that IVSd、LVPW and EF in rAAV-δ-SG-MSCs group significantly increased with LVDd and LVSd decreasing (P<0.05), while there was no difference between MSCs group and rAAV-δ-SG group. In rAAV-δ-SG-MSCs group, cardiomyocytes and cardiac structures were mature with more actin and blood vessels density detected by immunostaining (P<0.01), which was also confirmed by electron-microscope showing integrated cardiomyocytes and newborn blood vessels. Conclusion: Mesenchymal stem cells expressingδ-SG by rAAV can integrate into host cells. As compared with rAAV-δ-SG and MSCs , rAAV-δ-SG-MSCs transplantation can effectively repair myocardial sarcomere filament cytoskeletal protein, persistently repair TO-2 hamster myocardium, improve heart function, and delay the expansion of heart ventricle cavity..

关 键 词：δ-SG基因 骨髓间充质干细胞 心肌修复 心功能 细胞移植 

分 类 号：R542.2[医药卫生—心血管疾病]