目的:观察将脂肪间充质干细胞移植人犬心肌梗死模型后,犬心功能的变化情况。方法:实验于2005—08/2006-03在解放军总医院心内科实验室及解放军总医院实验动物中心完成,选用杂种犬13只,按随机数字表法分为2组,对照组7只,脂肪间充质干细胞移植组6只。①两组犬均进行心肌梗死模型制备,取犬脂肪间充质干细胞进行体外分离培养和DAPI标记,脂肪间充质干细胞移植组犬心外膜下共注射2×10^6个DAPI标记后的脂肪间充质干细胞,对照组注射相同体积IMDM完全培养基。常规关闭胸腔,继续饲养4周。②于细胞移植术前、术后4周行超声心动检查,测定左心室射血分数、左心室舒张末容积及左心室收缩末容积;术后4周行血流动力学检查,测定左心室舒张末压力、左心室收缩末压力、左心室室内压最大上升速率及左心室室内压最大下降速率。术后4周后处死动物取心脏标本观察移植细胞存活及分化情况。结果:对照组1只犬于移植术后第2天死亡,进入结果分析共12只犬,对照组和脂肪间充质干细胞移植组各6只。①移植术后4周移植细胞存活及分化的观察结果:在脂肪间充质干细胞移植组心肌组织冰冻切片中可见发出DAPI蓝色荧光染色的细胞核,同一视野下还可看到发绿色荧光的细胞浆。②移植术后4周两组犬心功能的比较:脂肪间充质干细胞移植组与对照组比较,左心室室内压最大下降速率、左心室舒张末压力、左心室舒张末容积及左心室收缩末容积减少(P〈0.05~0.01),左心室射血分数、左心室收缩末压力、左心室室内压最大上升速率增加(P〈0.05-0.01)。结论:脂肪间充质干细胞移植可在心肌细胞内存活,并可分化为心肌细胞,改善急性心肌梗死后心力衰竭犬的心脏功能。
AIM: To study the change of heart function of canines after the transplantation of adipose tissue-derived mesenchymal stem cells (ADMSCs) into canine models with myocardial infarction. METHODS: The experiment was performed at the Laboratory of Department of Cardiology and Experimental Animal Center, General Hospital of Chinese PLA from August 2005 to March 2006. Totally 13 hybrid canines were selected and randomly divided into 2 groups: control group with 7 canines and ADMSCs transplantation group with 6 canines. ① The canines in the two groups were established into myocardial infarction models. The ADMSCs isolated, cultured in vitro and marked by DAPI. Then 2×10^6 marked ADMSCs were transplanted into canines in the ADMSCs transplantation group through epidium. Those in the control group were treated with IMDM medium of the same volume. Thoracic cavity was closed routinely, and then cultured for 4 week successively. ②Before transplantation and 4 weeks after transplantation, all canines received ultrasonic cardiogram (UCG) test to determine left ventricular ejection fraction (LVEF), left ventricular end-diastolic volume (LVEDV) and left ventricular end-systolic volume (LVESV). Four weeks after transplantation, all canines received haemodynamics test to determine left ventricular end- diastolic pressure (LVEDP), left ventricular end-systolic pressure (LVESP) and +dp/dtmax and -dp/dtmax. Four weeks later, canines were killed to obtain heart sample, and then the transplanted cells survival and differentiation was observed. RESULTS: One canine in the control group died at day 2 after operation, and totally 12 canines were involved in the result analysis, six in the control group and six in the ADMSCs transplantation group. ①Observation of the transplanted cells survival and differentiation 4 weeks after transplantation: Nucleus with DAPI blue fluorescent staining appeared in frozen section of myocardial tissues in the ADMSCs transplantation group. Cytoplasm with green flu