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目的:研究骨髓间充质干细胞(mesenchymal stem cells,MSCs)移植到急性心肌梗死区心肌样细胞再生后ne-sprin蛋白的表达情况。方法:大鼠16只,随机分为实验组和对照组,每组8只。利用密度梯度分离法和贴壁培养MSCs,流式细胞仪检测MSCs表面相关抗原、DAPI标记MSCs备用;SD大鼠开胸手术结扎心脏冠脉左前降支建立心肌梗死模型,并在实验组梗死区注射MSCs,对照组注射DMEM;术后3周取梗死区心肌,利用免疫细胞学化学方法鉴定肌节肌动蛋白和肌钙蛋白的表达,免疫荧光技术和蛋白质印迹鉴定nesprin蛋白在MSCs移植后的表达。结果:MSCs移植到梗死缺血区3周后可以用DAPI示踪、并有心肌细胞特异蛋白呈阳性表达;免疫荧光和蛋白质印迹鉴定在MSCs移植前后有nesprin蛋白的表达,移植后出现nesprin蛋白表达量的增加。结论:MSCs在梗死心肌内移植具有分化再生的能力,nesprin蛋白在MSCs移植后出现表达量的增加,可能在MSCs分化为心肌细胞样过程中起到一定的作用。
OBJECTIVE: To study the expression of ne-sprin protein after mesenchymal stem cells (MSCs) transplanted into cardiomyocyte-like cells in acute myocardial infarction area. Methods: Sixteen rats were randomly divided into experimental group and control group, with 8 rats in each group. The MSCs were detected by density gradient centrifugation and adherent culture. The surface antigens of MSCs were detected by flow cytometry, and DAPI-labeled MSCs were prepared for backup. The myocardial infarction model was established by ligating left anterior descending coronary artery in the thoracotomy of SD rats. MSCs were injected into the control group and DMEM was injected into the control group. Myocardial infarction area was taken at 3 weeks after operation, and the expression of sarcomeric actin and troponin was identified by immunocytochemistry. The expression of nesprin protein was identified by immunofluorescence and Western blotting expression. Results: MSCs were transplanted into ischemic zone of infarction for 3 weeks and then could be traced by DAPI, and cardiomyocyte-specific protein was positive. Immunofluorescence and Western blotting showed that nesprin protein was expressed before and after MSCs transplantation. Increase in volume. CONCLUSION: MSCs have the ability to differentiate and regenerate in the infarcted myocardium. The increased expression of nesprin protein after MSCs transplantation may play a role in the differentiation of MSCs into cardiomyocyte-like process.