【摘要】 目的探讨自体骨髓间充质干细胞(mesenchymal stem cells,MSCs)经冠脉移植对猪急性心肌梗死后心功能的影响及其可能机制。方法 13头巴马香猪随机分为对照组和MSCs移植组,经髂后上棘抽取骨髓,密度梯度结合贴壁培养法分离、纯化、扩增MSCs。采用心导管介入球囊封堵猪冠状动脉左前降支(left anterior descending coronary ar-tery,LAD)建立急性心肌梗死模型。模型建立后MSCs移植组和对照组分别经OTW球囊于梗死相关冠脉注入4,6-二咪基-4-联苯基吲哚(4,6-Diamidino-2-Phenylindole,DAPI)标记过的MSCs和DMEM培养液。于制模前、细胞移植后4周行超声心动图检测左室内径和左室射血分数(left ventricular ejection fraction,LVEF)。不同时间点留取血标本检测血管内皮生长因子(vascular endothelial growth factor,VEGF)的浓度。移植后4周处死动物,TTC染色测量梗死面积,通过Ⅷ因子免疫组化染色比较两组微血管密度的差异。结果移植后4周,与对照组相比,MSCs移植组LVEF显著提高[(40.96±0.78)%vs.(34.54±1.55)%,P<0.05],TTC染色梗死面积明显减小[(18.0±5.8)%vs.(23.0±7.2)%,P<0.05],毛细血管计数明显增多[(103.6±4.4)/mm2 vs.(50.5±3.8)/mm2,P<0.05];MSCs移植组从术后第5天起VEGF表达较对照组增加,术后第7～10天达高峰,术后4周仍高于对照组(P<0.05)。结论经导管移植自体MSCs可以使血清VEGF含量增加,诱导血管新生,增加缺血区和梗死区血流灌注,改善心脏功能。更多还原

【Abstract】 Objective To investigate the effects of transplantation of autologous bone marrow mesenchymal stem cells via coronary artery by cardiac catheter on myocardial infarction(MI)and the action mechanisms.Methods Total 13 Bama swine were randomly divided into experimental group(n=7)and control group(n=6).Left anterior descending coronary arteries were occluded by balloon catheter to establish MI models.A total of 20 mL bone marrow was separated from the postero-superioriliac spine of models.Two mL MSCs and DMEM culture were respectively infused into the infarct-related coronary artery via an over-the-wire balloon.Cardiac functions were evaluated by ultrasonic cardiogram.The serum vascular endothelial growth factor(VEGF)levels were determined before AMI and after transplantation.At 4th week after transplantation,the size of MI was measured by using TTC dyeing.The vessels were counted by immunohistochemistry with factor Ⅷ monoclonal antibody.ResultsAs compared with control group,LVEF was significantly improved[(40.96±0.78)%vs.(34.54±1.55)%,P<0.05]and the sizes of MI were significantly decreased in experimental group [(18.0±5.8)%vs.(23.0±7.2)%,P<0.05].Immunohistochemistry revealed that the number of vessels stained with factor Ⅷ monoclonal antibody was greater than in control group [(103.6±4.4)/mm2vs.(50.5±3.8)/mm2,P<0.05].Serum levels of VEGF in experimental group were significantly elevated as compared with control group.Conclusion Transplantation of autologous MSCs via coronary artery by cardiac catheter can elevate the expression of VEGF,increase the amount of vessels,which may contribute to the improvement of cardiac functions.更多还原