同种异体骨髓间充质干细胞心内膜移植治疗小型猪急性心肌梗死

Transplantation of exogenous bone marrow mesenchymal stem cells by endocardium injection for treatment of acute myocardial infarction

目的：探讨同种异体骨髓间充质干细胞（MSCs）心内膜移植治疗小型猪急性心肌梗死的疗效。方法：分离、纯化、扩增培养小型猪MSCs;开胸结扎第1与第2对角支间前降支中段形成急性前壁心肌梗死的动物模型,心肌梗死后2-3周经心内膜注射移植DAPI标记的同种异体MSCs,心肌梗死前、后及移植后3个月行超声心动图及生化检查,并取梗死周围心肌组织冰冻切片及苏木精-伊红染色,免疫荧光法鉴定结蛋白（desmin）和心肌肌钙蛋白I（cTnI）的表达。结果：每40ml骨髓液经2-3周传3代平均获得（3.81±0.09）×107个细胞,移植后3个月左室射血分数较心肌梗死后及对照组改善明显,分别为（56.7±0.8）%,（40.1±1.2）%,（34.9±0.9）%（均P〈0.05）,肝肾功能、血糖及血气分析正常,3个月冰冻切片可见坏死心肌周围细胞核蓝染的移植细胞,免疫组化提示desmin及cTnI表达阳性。结论：同种异体MSCs经心内膜移植安全有效,心功能明显改善。

Objective：To evaluate the therapeutic effect of intramyocardial transplantation with exogenous bone marrow mesenchymal stem cells （MSCs） in the setting of acute myocardial infarction. Method： Acute myocardial infarction was induced by ligating the left anterior descending coronary artery （LAD） in swine. Mesenchymal stem cells （MSCs） were freshly isolated from the bone marrow blood by 1. 077 g/ml density-gradient centrifugation. 2 -3 weeks after cultured in the complete cell-culturing medium, the purified exogenous MSCs or no serum medium were transplanted into target ischemia endocardium by reconstructed injection catheter. LV function was estimated by echocardiogram and blood biochemistry after 3 months transplantation. Desmin and cardiac-specific cardiac troponin Ⅰ （cTnⅠ） was examined with immunohischemistry method around infracted region after 3 months transplantation. Result：MSCs were harvested （3.81±0.09）×10^7 ceils after cultured three generations over 2-3 weeks. DAPI-labeled MSCs in 1ml medium and medium only were multi-sites injected into the ischemia I.AD territory with injection catheter. Left ventricular ejection fractions were （ 56.7 ± 0.8） % , （ 40.1±1.2 ） %, （ 34.9 ± 0.9） % （P〈0.05） at 3 months in transplantation group, control group and post AMI respectively. The function of liver and kidney, blood glucose, artery blood gas analysis were normal after transplan tation. The positive stains of transplanted cells, desmin and cTnⅠ were found in infarct myocardium at three month after transplantation. Conclusion：The therapy of generated exogenous MSCs transplantation is safe and effective.