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目的:观察经冠状动脉注射成年犬自体骨髓基质干细胞对梗死后心功能的影响。方法:本实验于2001-05/2002-04在解放军第二军医大学附属长征医院心内科实验室完成。选用成年犬8只,随机分为细胞移植组5只,对照组3只。结扎犬冠状动脉前降支制备心肌梗死模型;细胞移植组将体外培养、诱导的自体骨髓基质干细胞标记后在心肌梗死后4周经冠状动脉植入心脏;对照组植入等量培养基。胸内心脏超声检查测定左冠状动脉血流阻断前,再灌注后30min及移植后4周左心室射血分数;胸外心脏超声检查测定左冠状动脉血流阻断前、移植前、移植后4周左心室射血分数。用成组t检验和随机区组方差分析完成统计学分析。结果:进入结果分析犬8只。①植入的骨髓基质干细胞在移植犬心肌组织内存活情况:细胞移植组免疫组织化学检查结蛋白及肌钙蛋白I染色阳性,表示已生成心肌样细胞。对照组心脏标本中可见瘢痕组织;心肌细胞结蛋白染色阴性,肌钙蛋白I染色阳性。②移植犬左室功能的左室射血分数:经胸外心脏彩色多普勒超声检查提示,细胞移植组细胞移植前明显低于血流阻断前和移植后4周犤(54.86±7.12)%,(61.78±5.37)%,(61.74±2.93)%,P<0.01犦;细胞移植组细胞移植后4周接近血流阻断前(P>0.05);对照组移植后4周和移植前明显低于血流阻断前(P<0.01)。③移植犬左室功能的左室射血分数:经胸内心脏彩色多普勒超声检查反映,细胞移植组再灌注后30min明显低于血流阻断前和移植后4周犤(49.98±6.79)%;(61.56±3.71)%,(64.56±5.27)%,P<0.01犦。移植后4周明显高于血流阻断前(P<0.05)。对照组移植后4周和再灌注后30min明显低于血流阻断前(P<0.05,0.01)。结论:成年心肌梗死犬自体骨髓基质干细胞经冠状动脉移植后,在心肌组织内存活并分化为肌源性细胞使左心室功能改善。
Objective: To observe the effects of autologous bone marrow stromal stem cells injected into coronary artery on the cardiac function after infarction in coronary artery. Methods: The experiment was performed at the Cardiology Laboratory of Changzheng Hospital affiliated to the Second Military Medical University of Chinese PLA from May 2001 to April 2002. 8 adult dogs were randomly divided into five cell transplantation groups and three control groups. The model of myocardial infarction was prepared by ligating the anterior descending branches of canine. The cell transplantation group was cultured in vitro, and the induced autologous bone marrow stromal stem cells were labeled and then implanted into the heart by coronary artery 4 weeks after myocardial infarction. The control group was implanted with equal volume of medium. Intrathoracic echocardiography was used to measure the left ventricular ejection fraction before occlusion of left coronary artery, 30 min after reperfusion and 4 weeks after transplantation. The left ventricular ejection fraction was measured before and after left coronary artery occlusion by thoracic echocardiography 4 weeks left ventricular ejection fraction. Statistical analysis was performed using a group t-test and randomized block analysis of variance. Results: Into the results of analysis of dogs 8. ① Survival of implanted bone marrow stromal stem cells in myocardial tissue of transplant dogs: Immunohistochemical staining of desmin and troponin I in the cell transplantation group showed that cardiomyocyte-like cells were formed. In the control group, scar tissue was found in the cardiac specimens; the myocardial cells were negative for staining for desmin and positive for troponin I staining. Left ventricular ejection fraction of transplanted dogs with left ventricular function: The extracorporeal color Doppler echocardiography showed that the cell transplantation group was significantly lower than that before transplantation and 4 weeks after transplantation (54.86 ± 7.12) %, (61.78 ± 5.37)% and (61.74 ± 2.93)% respectively, P <0.01 犦. The cells in the cell transplantation group were close to the blood flow 4 weeks after the transplantation (P> 0.05) Significantly lower than before blood flow was blocked (P <0.01). Left ventricular ejection fraction of transplanted canine left ventricular function: Color Doppler ultrasound examination of intrathoracic heart showed that the cell transplantation group 30min after reperfusion was significantly lower than that before blood flow block and 4 weeks after transplantation 犤 (49.98 ± 6.79 );%; (61.56 ± 3.71)%, (64.56 ± 5.27)%, P <0.01 犦. 4 weeks after transplantation was significantly higher than before blood flow blocking (P <0.05). The control group 4 weeks after transplantation and 30 min after reperfusion were significantly lower than those before blood flow block (P <0.05, 0.01). CONCLUSION: Autologous bone marrow stromal stem cells from adult myocardial infarction dogs survive in the myocardium and differentiate into myogenic cells to improve left ventricular function after coronary artery transplantation.