非人工通气下建立大鼠心肌内干细胞移植模型

小动物心肌内干细胞移植术较多采用气管插管呼吸机辅助通气和剪断胸骨或肋骨而开胸造模，其损伤大、操作复杂、且影响模型存活率。 目的：在非人工通气条件下，以较小创伤，简单快速建立正常及心肌梗死大鼠心肌内干细胞移植模型。 方法：将SD大鼠随机均分2组：心肌梗死移植组建立心肌梗死模型后接受骨髓间充质干细胞移植，在心肌梗死周边区分5点迅速注射骨髓间充质干细胞；正常移植组于左心室前壁心外膜下进行注射，方法及注射量同心肌梗死移植组。移植后4 d，取大鼠心脏行冰冻切片，荧光显微镜观察绿色荧光蛋白标记的骨髓间充质干细胞存活与分布，免疫荧光染色检测骨髓间充质干细胞心肌特异性蛋白的表达。 结果与结论：心肌梗死移植组模型存活率为80%，正常移植组存活率为93%。移植后4 d，移植区可见大量骨髓间充质干细胞，呈带状分布，排列方向与心肌纤维一致，并表达缝隙连接蛋白Connexin43和肌球蛋白重链MYH。结果提示：在非人工通气条件下，快速、简单、有效地建立了大鼠心肌内干细胞移植模型。

Establishing a rat model of intramyocardial stem cell transplantation without artificial ventilation

BACKGROUND: Models of intramyocardial transplantation of stem cells in small experimental animals are often established with artificial ventilation by endotracheal intubation, in addition to cutting sternum or ribs. The operation is seriously wounded and complicated, which may impact on survival of models. OBJECTIVE: To establish a model of intramyocardial transplantation of stem cells in normal and infarcted rats without artificial ventilation. METHODS: Sprague Dawley rats were assigned to two groups. Myocardial infarction group: rats, following successful establishment of myocardial infarction model, received bone marrow mesenchymal stem cells (BMSCs) transplantation. For each rat, BMSCs were injected into surrounding sites of the infracted region by five points. In normal group, BMSCs with the same quantity were injected into epicardial sites in the left ventricle anterior wall by the same method. Rats in each group were killed 4 days after transplantation, and hearts were obtained for frozen sections. The distribution and survival of BMSCs labeled with green fluorescent protein (GFP) were observed under fluorescence microscope. Immunofluorescence staining was used to examine the cardiac-specific proteins expression on BMSCs. RESULTS AND CONCLUSION: The survival rate was 80% in transplantation model in myocardial infarction group, and 93% in normal group. 4 days after transplantation, a lot of BMSCs, which showed zonal distribution with arranging in parallel with myocardial fibers, could be observed in the transplantation area. Furthermore, Connexin43 and myosin heavy chain MYH were also detected on BMSCs. Results indicate that using this method, a model of intramyocardial transplantation of stem cells in rat can be established simply and effectively without using artificial ventilation.