非诱导同种异体骨髓基质细胞心肌移植后细胞增殖分化的结局

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。

Outcomes of cell proliferation and differentiation after myocardium transplantation of uninduced allogeneic bone marrow stromal cells

BACKGROUND: The studies about cellular cardiomyoplasty (CCM) of bone marrow stromal cell (BMSC) are centered on modeling autologous cell transplantation while the study of myocardium transplantation of allogeneic BMSC is seldom reported domestically.OBJECTIVE: To study the hypothesis that the allogeneic BMSC, after transplanted into the myocardial infarction (MI) regions, can survive, further proliferate, differentiate, and its effects on host hearts.DESIGN: A randomized and controlled trial with experimental animals as subjects.SETTING: Laboratory of Internal Cardiology, Affiliated Union Hospital, Tongji Medical College, Huazhong University of Science and Technology.MATERIALS: Ten Wistar rats of one-month old, body mass of 100-120 g and unconfined sex were used to culture BMSC while eighty female Wister rats of three-month old, body mass of 200-250 g were used for animal models.METHODS: The experiment was completed in the Laboratory of of Internal Cardiology, Affiliated Union Hospital, Tongji Medical College,Huazhong University of Science and Technology from June to December 2004. ①Eighty rats were used to establish the acute myocardial infarction (AMI) models by ligating the left anterior descending branch of coronary artery, and 45 of them were successful. ② After 4 weeks, the models were divided into 2 groups at random. In the experiment group (n=25), the passaged and uninduced BMSC cultured in vitro were injected into the MI region of the recipients while only medium was injected in the control group (n=20). ③At 4 weeks after implantation,the hemodynamic indexes of recipients' hearts were examined. Then the samples were obtained to detect the survival, differentiation and angiogenesis status of the removal cells.MAIN OUTCOME MEASURES: ①Comparison of hemodynamic indexes in rats of two groups②Results of the implantation cells③Changes of angiogenesis in rats of two groups.RESULTS: Totally 13 rats in the experiment group and 11 in the control group were involved in the result analysis. ①The hemodynamic indexes of rats were significantly improved in the experiment groups compared with the control group Left ventricle systolic blood pressure (LVSBP): (88.61±5.99),(76.93±4.75) mm Hg, left ventricle end-diastolic pressure(LVEDP): (7.72±1.36),(12.77±2.76) mm Hg, P ＜ 0.05;The maximum changing velocity of LVSBP:(2 365.26±266.31),(2 025.04±230.25) mm Hg/s; The maximum changing velocity of LVDBP:(2 313.26±159.30),(2 140.12±191.03) mm Hg/s]. ②After implanted in the MI region, the allogeneic BMSC passed the acute inflammation period and did not induce the remarkable reject reaction of transplantation. The BMSC in the infarcted region were mainly differentiated into fibroblast. Some cells around the infarcted region were differentiated into endothelial cells, and improved the angiogenesis. ③The number of angiogenesis in and around the transplantation regions was significantly higher in the experiment group than in the control group (P ＜ 0.01).CONCLUSION: The allogeneic BMSC can not form cardiomyocyte in the infarcted region after cell transplantation, and the engendered endothelial cells of blood vessels may promote the angiogenesis after AMI and ameliorate the cardiac function.