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目的探讨脂肪间充质干细胞(adipose tissue-derived mesenchymal stem cells,ADMSCs)移植对兔心肌梗死后微血管生成的可能机制及心功能的变化情况。方法 30只健康日本大耳白兔,完全随机分为假手术组、心肌梗死对照组和ADMSCs移植组(ADMSCs组)。结扎兔前室间支,建立急性心肌梗死(acute myocardial infarction,AMI)动物模型,AMI1h内将4’,6-二脒-2-苯基吲哚(4’,6-diamidio-2-phenylindole,DAPI)标记的第3代ADMSCs植入ADMSCs组梗死心肌,对照组及假手术组注射等量磷酸盐缓冲溶液(phosphate buffered solution,PBS)。术前及术后4周分别做超声心动图检查其心功能变化。处死动物,取其心脏,采用免疫组织化学法检测梗死区新生血管微血管密度,RT-PCR检测血管内皮生长因子(vascular endothelial growth factor,VEGF)和碱性成纤维生长因子(basic fibroblast growth factor,bFGF)的表达。结果 ADMSCs组荧光显微镜下可以观察到DAPI标记细胞存在,部分分化为心肌样细胞;超声心动图证实移植后4周ADMSCs组心功能较心肌梗死对照组改善(P<0.05);ADMSCs组较心肌梗死对照组梗死区微血管密度明显增高[(20.00±2.65)vs(7.75±2.12),P<0.05];ADMSCs组bFGF和VEGF的表达水平均较心肌梗死对照组明显增高[(0.590±0.028)vs(0.569±0.021),(0.913±0.030)vs(0.886±0.049),P<0.05]。结论ADMSCs移植促进心肌梗死区微血管的生成,并改善心脏功能,其机制可能与促进bFGF、VEGF的表达有关。
Objective To investigate the possible mechanism and cardiac function of adipose tissue-derived mesenchymal stem cells (ADMSCs) implanted into rabbit after myocardial infarction. Methods Thirty healthy Japanese white rabbits were randomly divided into sham operation group, myocardial infarction control group and ADMSCs transplantation group (ADMSCs group). The anterior branch of the rabbit was ligated and the animal model of acute myocardial infarction (AMI) was established. In the AMI1h, 4 ’, 6-diamidio-2-phenylindole DAPI labeled 3rd generation ADMSCs were implanted into the infarcted myocardium of ADMSCs group. The control group and the sham operation group were injected with the same amount of phosphate buffered solution (PBS). Echocardiography was performed preoperatively and 4 weeks after surgery to examine changes in cardiac function. The animals were sacrificed and the heart was taken. The density of neovascular microvessels in the infarct area was detected by immunohistochemistry. The expressions of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) )expression. Results The DAPI-labeled cells were observed under fluorescence microscope in ADMSCs group, and some differentiated into cardiomyocyte-like cells under echocardiography. The cardiac function of ADMSCs group was improved 4 weeks after transplantation compared with myocardial infarction control group (P <0.05) (20.00 ± 2.65) vs (7.75 ± 2.12), P <0.05]. The expression of bFGF and VEGF in ADMSCs was significantly higher than that in myocardial infarction control group [(0.590 ± 0.028) vs ( 0.569 ± 0.021), (0.913 ± 0.030) vs (0.886 ± 0.049), P <0.05]. Conclusion Transplantation of ADMSCs can promote angiogenesis and improve cardiac function in myocardial infarction area. Its mechanism may be related to promoting the expression of bFGF and VEGF.