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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 observe the effect of bone marrow-derived mesenchymal stem cells (MSCs), 5-azacytidine (5-Aza) -induced cardiomyocyte-like cells and 2 kinds of cells combined transplantation on functional aneurysms after myocardial infarction in rats. Methods: Rat MSCs were cultured in vitro and cardiomyocytes induced by 5-Aza. Left anterior descending coronary artery was ligated to form a myocardial infarction. Four weeks later, echocardiography was used to detect and screen functional aneurysms. The patients were divided into 4 groups: group A (n = 10) Peripherally injected MSCs (106-107), B group (n = 10), cardiomyocytes (106-107), C group (n = 10) 107). Group D (n = 10) was the control group with 0.9% sodium chloride injection. Four weeks after operation, the cardiac function of rats was measured by echocardiography and hemodynamics. In addition, histological evaluation of capillary density after cell transplantation, Masson’s trichrome staining method to measure the extent of aneurysm. Results: Compared with D group, left ventricular diastolic diameter and left ventricular systolic diameter were significantly reduced in group A, B and C (P <0.05), short axis shortening and left ventricular ejection fraction (P <0.05), and C group was superior to A group and B group (P <0.05). There was no significant difference between A group and B group (P> 0.05). Compared with group D, group D, group A, group B and group C showed a significant increase in left ventricular systolic and diastolic pressure and maximum positive and negative rates of left ventricular (P <0.05), and increased in group C (P <0.05). There was no significant difference between group A and group B (P> 0.05). The hematoxylin-eosin staining showed that the capillary density in group A (3.452 ± 0.168 / high power field) and group C (3.383 ± 0.129 / high power field) were higher than those in group D (1.827 ± 0.052 / high power field) P <0.05). There was no significant difference between group B (1.917 ± 0.038 / high power field) and group D (P> 0.05). The results of Masson ’s trichrome staining in evaluating the extent of ventricular aneurysm showed that in group A (21.32 ± 0.90)%, in group B (22.14 ± 0.74)% and in group C (21.98 ± 0.51)% The extent of aneurysm was smaller in group D than in group D [(25.70 ± 1.71)%], the difference was statistically significant (P <0.05). CONCLUSION: MSCs transplantation, cardiomyocyte-like cell transplantation, and combined transplantation of both have a repair effect on the rat aneurysm. The combination of MSCs and cardiomyocyte-like cells improves heart function and ventricular remodeling better than single cell transplantation.