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目的探讨脐血间质干细胞移植对梗死心肌残存心肌细胞凋亡的影响。方法无菌条件下采集成年杂种妊娠犬脐血,体外分离、纯化、培养、扩增脐血间质干细胞,经5-氮杂胞苷(5-aza)体外诱导向肌源性细胞分化,DAPI荧光标记,经左冠状动脉前降支灌注移植入成年杂种犬急性心肌梗死模型区域,2、4、8周取心肌标本,采用HE染色及TUNEL法检测犬心肌梗死模型中脐血间质干细胞移植后心肌细胞的凋亡情况。结果免疫组化法检测结果表明,培养的细胞为脐血间质干细胞,在5-aza诱导作用下可向肌源性细胞定向分化,2、4、8周后在移植区域观察到带荧光的心肌纤维和细胞核,排列方向与残留心肌组织基本一致,荧光标记的肌纤维相互连接。经TUNEL法检测可见在脐血间质干细胞移植组和对照组心肌梗死区域均有凋亡的心肌细胞分布,但移植组凋亡细胞明显低于非移植组;HE染色有相似的阳性率,阳性细胞多具有凋亡的形态特征。结论脐血间质干细胞移植梗死心肌除能形成心肌再生外,尚能减少心肌梗死后梗死区域的心肌细胞凋亡,达到延缓心室重构的目的。应用TUNEL法检测心肌细胞凋亡有较强的实用性。
Objective To investigate the effect of umbilical cord blood mesenchymal stem cell transplantation on myocardial cell apoptosis in infarcted myocardium. Methods Adult umbilical cord blood of pregnant dogs were collected under aseptic conditions, isolated, purified, cultured and expanded umbilical cord blood mesenchymal stem cells, induced by 5-azacytidine (5-aza) into myogenic cells, DAPI Fluorescently labeled, the left anterior descending coronary artery perfusion perfusion into adult mongrel dogs acute myocardial infarction model area, 2, 4, 8 weeks to take myocardial samples, HE staining and TUNEL assay canine myocardial infarction model of umbilical cord blood mesenchymal stem cell transplantation After myocardial cell apoptosis. Results The results of immunohistochemistry showed that the cultured cells were umbilical cord blood mesenchymal stem cells which could differentiate into myogenic cells under the induction of 5-aza. After 2, 4 and 8 weeks, the cells with fluorescence Myocardial fibers and nuclei, arranged in the direction of the residual myocardial tissue is basically the same, fluorescent labeled muscle fibers connected to each other. TUNEL assay showed that the apoptotic cardiomyocytes were distributed in the infarcted area of cord blood MSCs and the control group, but the apoptotic cells in the transplantation group were significantly lower than those in the non-transplantation group; the positive rates were similar in HE staining More cells have the morphological characteristics of apoptosis. Conclusion In addition to the myocardial regeneration induced by myocardial infarction in umbilical cord blood mesenchymal stem cells transplantation, myocardial infarction can be reduced myocardial infarction area myocardial infarction, to delay the purpose of ventricular remodeling. TUNEL method used to detect myocardial apoptosis has strong practicality.