摘要
目的初步观察自体骨髓间充质干细胞(MSCs)移植后在心梗区存活并分化为心肌细胞的情况,探讨MSCs经冠脉移植对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MSCs移植组(n=7)和DMEM培养液对照组(n=6)。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。通过心导管介入球囊封堵冠状动脉左前降支(LAD)第2对角支以远建立急性心肌梗死模型。模型建立后即刻,细胞移植组和对照组分别经梗死相关冠脉注入4,6-二咪基-4-联苯基吲哚(DAPI)标记过的MSCs和DMEM培养液。于心梗前、细胞移植后4周对猪进行超声心动图检查。移植后4周处死动物,观察心脏大体结构,苏木精-伊红染色及Masson三色染色法观察显微结构改变。通过免疫组化检测移植细胞的肌钙蛋白T(Cardiac Troponin T,c-TnT)、缝隙连接蛋白43(Connexin43,Cx-43)的表达。结果移植后4周,MSCs移植组和对照组比较射血分数(LVEF)显著提高(P<0.05),左室收缩末期内径(LVESD)和左室舒张末期内径(LVEDD)显著减小(P<0.05)。苏木精-伊红染色发现对照组病变区心肌变性、坏死,大量炎性细胞浸润,梗死区与非梗死区界限清楚,移植组梗死区与非梗死区界限不清,且在梗死区中可见部分排列相对整齐的肌纤维结构。Masson染色显示移植组梗死区和交界区内胶原纤维融合较少,排列较为有序,而对照组梗死区和交界区内胶原纤维斑块状融合明显,排列紊乱。MSCs移植组DAPI阳性细胞c-TnT、Cx-43表达阳性。结论在急性心肌梗死造模后即刻进行经冠状动脉MSCs自体移植,MSCs可以在梗死局部存活并向心肌细胞分化,从而抑制左室重构,显著改善心功能。
Objective To observe the survival and differentiation of cardiomyocytes after the transplantation of autologus bone marrow mesenchymal stem cells（MSCs）,and discuss the influence of MSCs transplantation via coronary artery by cardiac catheter on the cardiac function following acute myocardial infarction（AMI）.Methods Thirteen Bama swines were randomly divided into MSCs group（n=7）and DMEM culture medium control group（n=6）.MSCs were isolated from the posterior superior iliac spine and cultured in vitro.These pigs underwent occlusion of the left anterior descending coronary artery（LAD）to establish AMI models.MSCs labeled with DAPI and DMEM culture medium were respectively infused into the infarct-related coronary artery via an over-the-wire balloon.Cardiac function was evaluated prior to AMI and 4 weeks after transplantation by echocardiography.HE staining and Masson staining were used to observe the microstructure.Immunohistochemistry was used to detect c-TnT and connexin-43（Cx-43）4 weeks after transplantation.Results Four weeks after MSCs transplantation,left ventricular ejection fraction（LVEF） was improved significantly（P0.05）,and left ventricular end-systolic dimension（LVESD）and left ventricular end-diastolic dimension（LVEDD）were decreased significantly（P0.05） as compared with control group.HE and Masson staining showed that there were severe fibrosis and inflammatory cell infiltration in control group,with seldom surviving myocardium in infracted zone.However,there were less fibrosis and inflammatory cell infiltration in MSCs group.Immunohistochemical examination revealed that the DAPI positive cells also expressed c-TnT and Cx-43 in MSCs group.Conclusion MSCs transplanted via the coronary artery can survive in the infracted scar and differentiate into cardiomyocytes,which could improve the condition of left ventricular remodeling and recover the cardiac functions.
出处
《华中科技大学学报（医学版）》
CAS
CSCD
北大核心
2012年第5期562-565,共4页
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
基金
河南省教育厅科研课题(No.2004922073)
河南省科技厅科研课题(No.0524410058)
关键词
骨髓间充质干细胞
细胞移植
急性心肌梗死
心功能
bone marrow mesenchymal stem cells
cell transplantation
acute myocardial infarction
cardiac function
