多种细胞移植治疗大鼠心肌梗死后功能性室壁瘤的疗效比较

Comparative investigation of different cells transplantation on cardiac function of rats with ventricular aneurysm after myocardial infarction

目的:观察比较骨髓间充质干细胞(MSCs)、5-氮杂胞苷(5-Aza)诱导分化的心肌样细胞及2种细胞联合移植治疗大鼠心肌梗死后功能性室壁瘤的疗效。方法:体外培养大鼠的MSCs,及用5-Aza诱导成的心肌样细胞。结扎大鼠冠状动脉左前降支,形成心肌梗死,4周后用超声心动图检测并筛选形成功能性室壁瘤者,分4组:A组(n=10),在室壁瘤瘤部及周边部点状注射MSCs(106～107个);B组(n=10),注射心肌样细胞(106～107个);C组(n=10),联合移植MSCs和心肌样细胞(106～107个);D组(n=10)为对照组,注射0.9%氯化钠。术后4周用超声心动图和血流动力学方法测定大鼠的心功能。另外,用组织学方法评价细胞移植后毛细血管密度,Masson氏三色染色法测量室壁瘤范围。结果:心脏彩超结果显示,与D组相比,A组、B组和C组的左室舒张内径、左室收缩内径均明显缩小(P<0.05),短轴缩短率及左室射血分数明显增大(P<0.05),且C组优于A组和B组(P<0.05),A组和B组比较,差异无统计学意义(P>0.05)。血流动力学检测结果显示,与D组相比,A组、B组和C组的左室舒缩压差和左室正负最大变化速率均明显增高(P<0.05),其中C组增高最明显(P<0.05),A组和B组比较,差异无统计学意义(P>0.05)。苏木精-伊红染色血管计数结果显示,A组(3.452±0.168/高倍视野)和C组(3.383±0.129/高倍视野)的毛细血管密度高于D组(1.827±0.052/高倍视野)(P<0.05),B组(1.917±0.038/高倍视野)与D组比较,差异无统计学意义(P>0.05)。Masson氏三色染色法评价室壁瘤范围(%)结果显示,A组[(21.32±0.90)%]、B组[(22.14±0.74)%]和C组[(21.98±0.51)%]的室壁瘤范围小于D组[(25.70±1.71)%],差异有统计学意义(P<0.05)。结论:MSCs移植,心肌样细胞移植,及二者联合移植对大鼠室壁瘤均有修复作用,联合MSCs和心肌样细胞移植改善心功能及心室重构方面效果优于单项细胞移植。

Objective：To investigate the therapeutic effect of mesenchymal stem cells （MSCs）transplantation combined with cardiomyocytes-like cells induced from MSCs on cardiac function of rats with functional ventricular aneurysm after myocardial infaretion. Methods：Isolated MSCs from rats bone marrow,and cultured in cell culture medium with 5-azacytidine（5 Aza）. Functional ventricular aneurysm model of rat was created by ligating the left enter/or descending coronary artery,and selected by ultrasonic cardiography（UCG） after 4 weeks. Rats with functional ventricular aneurysm were divided four groups： group A, MSCs（10^6- 10^7） were injected several differernt points into the broad area of ventricular aneurysm; group B, cardiomyocytes-like cells（10^6- 10^7） were implant; group C, MSCs transplantation combined with cardiomyocytes-like cells（10^6- 10^7）; group D was control group, sodium chloride were injected. Four weeks after transplantation, the cardiac function of the rats was examined by UCG and multichannel physiological recorder, capillary density was evaluated by histological method, and ventricular aneurysm size was measured with Masson＇s trichrome. Results： UCG showed that left ventricular end diastolic dimension（LVEDD）, left ventricular end systolic dimension （LVESD）of group A, B and C obviousy reduced compared with group D（P〈0. 05） ,and fractional shortening（△FS%） and left ventricular ejection fraction（LVEF% ） in three groups improved markedly compared with .group D（P〈0.05）, furthermore group C is better than groupA and B（P〈0. 05）,and there was no significant difference between groups A and B（P〉0. 05）. Multicbannel physi ological recorder showed that left ventricular developed pressure（LVDP）and （＋dp/dtmax） of groups A, B and C improved obviousy compared with group D（P〈0.05） ,C group was highest（P〈0.05）, and there was no significant difference between groups A and B（P〉0.05）. Result of HE showed that capillary density of group A（3. 452 ±0. 168/highpower field）and group C（3. 383±0. 129/ highpower field）were higher than group D（1. 827±0. 052/ highpower field）（P〈0.05）, there was no significant difference between group B（1. 917±0. 038/ highpower field） and group D（P〉0. 05）. Ventricular aneurysm size of group A（[21.32±0.90]% , group B[22.14±0.74]%） and group C（[21.98±0.51]%） markedly reduced as compared with group D（P〈0.05）. Conclusion：MSCs,cardiomyocytes-like ceils or combination cell transplantation have an active effect for functional ventricular aneurysm, however, combination cell transplantation could obviously improve the heart function and ameliorate ventricular remodeling.