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目的筛选骨髓间充质干细胞(mesenchymalstemcellsMSCs)移植改善心肌病大鼠心功能的相关基因,初步分析相关基因在MSCs改善心功能中的作用。方法用贴壁筛选法分离、扩增大鼠MSCs,并移植入经阿霉素腹腔注射构建的大鼠心肌病模型中。然后利用抑制性消减杂交技术(supressionsubtractivehybridization,SSH)筛选移植MSCs后大鼠心肌组织差异表达的相关基因,并进行分析。结果成功建立了心肌病大鼠模型,并进行了MSCs心肌移植,筛选出移植MSCs后心肌细胞表达上调的基因12条、表达下调的基因4条。经基因的序列检索分析显示,MSCs移植入大鼠心肌后与线粒体合成有关的基因和与心肌的收缩蛋白合成有关的基因表达上调;表达下调的基因分别为肌小节的线粒体肌氨酸激酶基因;核糖体的磷蛋白P2基因;α-晶状体蛋白基因;NADH-辅酶Q氧化还原酶Fe-S蛋白7基因。结论MSCs移植后可能通过刺激与能量合成相关和心肌收缩相关基因的表达,一方面促进心肌能量合成,另一方面MSCs可能分化为新的心肌细胞,增加心肌收缩力,从而改善心肌组织功能。
Objective To screen the genes related to cardiac function of cardiomyopathy rats after transplantation of mesenchymal stem cells (MSCs), and to analyze the role of related genes in improving cardiac function. Methods The rat MSCs were isolated and expanded by adherent screening method and were transplanted into rat model of cardiomyopathy induced by adriamycin intraperitoneal injection. Then, the related genes differentially expressed in rat myocardial tissue after MSCs transplanting were screened by SSH and analyzed. Results Cardiac myopathy rat model was successfully established and myocardial transplantation of MSCs was performed. Twelve genes were up-regulated in cardiomyocytes and four genes were down-regulated after transplanted MSCs. Genomic sequence analysis showed that the expression of genes related to mitochondrial synthesis and myocardial contractile protein synthesis was up-regulated after MSCs were transplanted into rat myocardium. The down-regulated genes were mitochondrial sarcosine kinase genes in muscle segments, The ribosomal phosphoprotein P2 gene; the α-crystallin gene; and the NADH-Coenzyme Q oxidoreductase Fe-S protein 7 gene. Conclusion MSCs may stimulate the synthesis of myocardial energy by stimulating the synthesis of energy-related genes and myocardial contraction, and on the other hand, MSCs may differentiate into new cardiomyocytes, increase myocardial contractility and improve myocardial tissue function.