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背景:关于骨髓基质细胞心肌成形的研究,目前都集中在模拟自体细胞移植方面。而同种异体骨髓基质细胞心肌移植的研究,国内报道较少。目的:观察大鼠同种异体骨髓基质细胞移植到心肌梗死区后能否存活,并进一步增殖、分化,以及对宿主心脏的影响。设计:随机对照动物实验。单位:华中科技大学同济医学院附属协和医院心血管内科实验室。材料:1月龄Wistar大鼠10只,雌雄不拘,体质量100～120g,用于取材培养骨髓基质细胞。3月龄雌性Wistar大鼠80只,体质量200～250g,用于制造动物模型。方法:实验于2004-06/12在华中科技大学同济医学院附属协和医院心血管内科实验室完成。①80只大鼠结扎冠状动脉左前降支制作急性心肌梗死模型,45只大鼠制作模型成功。②4周后,取传两代非经诱导的体外培养的同种异体大鼠骨髓基质细胞,注射到大鼠心肌梗死区,为移植组(25只)。同时设置注射培养基的对照组(20只)。③移植4周后检测受体心脏的血流动力学指标,然后取标本,检测移植细胞存活、分化和组织的血管新生状况。主要观察指标:①两组大鼠血流动力学检测结果比较。②移植细胞的结局。③两组大鼠血管新生的改变。结果:细胞移植组13只,对照组11只大鼠进入结果分析。①移植组大鼠心脏血流动力学指标较对照组明显改善犤左心室收缩压:(88.61±5.99),(76.93±4.75)mmHg,左心室舒张末压:(7.72±1.36),(12.77±2.76)mmHg,P均<0.05;左心室收缩压最大变化速率:(2365.26±266.31),(2025.04±230.25)(mmHg/s),左心室舒张压最大变化速率:(2313.26±159.30),(2140.12±191.03)mmHg/s犦。②同种异体骨髓基质细胞移植入心肌梗死区后能够度过急性炎症期而且不引起明显移植排斥反应;位于梗死区的移植细胞主要分化为成纤维细胞,部分位于心肌梗死区周围的细胞分化为血管内皮细胞,并促进了血管新生。③移植组心肌梗死区及其周围新生血管数目较对照组明显增加(P<0.01)。结论:同种异体骨髓基质细胞移植后未能在梗死区形成心肌样细胞,但所形成的血管内皮细胞产生了促进心肌梗死后血管新生、改善心功能的作用。
BACKGROUND: Studies on cardiac myogenic formation of bone marrow stromal cells have so far focused on the simulation of autologous cell transplantation. However, the study of allogeneic bone marrow stromal cells transplantation in the heart, less reported in China. Objective: To observe whether allograft marrow stromal cells of rats can survive after transplantation into myocardial infarction area, and further proliferation, differentiation, and the impact on the host heart. Design: Randomized controlled animal experiments. SETTING: Department of Cardiology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology. MATERIALS: Ten Wistar rats aged 1 month were randomly divided into male and female. The body weight was 100 ~ 120g, which was used to culture bone marrow stromal cells. Eighty-three-month-old female Wistar rats weighing 200-250 g were used to make animal models. Methods: The experiment was performed at the Department of Cardiology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology from June to December 2004. ①80 rats were ligated with left anterior descending artery of coronary artery to make acute myocardial infarction model, and 45 rats were made successfully. After4weeks, allograft bone marrow stromal cells of non-induced in vitro culture of two passages were taken and injected into the myocardial infarction area of rats for transplantation group (25 rats). At the same time set the control group of injection medium (20). ③ After 4 weeks of transplantation, the hemodynamics of the recipient’s heart were detected, and then specimens were collected to detect the engraftment, differentiation and tissue angiogenesis of the transplanted cells. MAIN OUTCOME MEASURES: ① Comparison of hemodynamic test results between the two groups. ② the outcome of transplanted cells. ③ angiogenesis changes in both groups. Results: 13 cells in the cell transplantation group and 11 rats in the control group were involved in the result analysis. ① Compared with the control group, the hemodynamics indexes of the left ventricle in the transplantation group were significantly improved: (88.61 ± 5.99), (76.93 ± 4.75) mmHg, left ventricular end-diastolic pressure: (7.72 ± 1.36), (12.77 ± 2.76) mmHg, both P <0.05. The maximum rate of change of left ventricular systolic pressure was (2365.26 ± 266.31), (2025.04 ± 230.25) mmHg / s, the maximum rate of change of left ventricular diastolic pressure was (2313.26 ± 159.30), (2140.12 ± 191.03) mmHg / s 犦. ② Allogeneic bone marrow stromal cells were able to survive the acute inflammatory phase after transplantation into myocardial infarction area without causing significant graft rejection. The transplanted cells located in the infarct area mainly differentiated into fibroblasts, and some of the cells located around the myocardial infarction area differentiated into Vascular endothelial cells, and promote angiogenesis. ③ The number of neovascularization in myocardial infarction area and its surroundings in transplantation group was significantly higher than that in control group (P <0.01). Conclusion: Allogeneic bone marrow stromal cells failed to form cardiomyocytes in the infarct zone after transplantation. However, the endothelial cells formed had the effect of promoting angiogenesis and improving cardiac function after myocardial infarction.