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目的:通过筛选差异表达miRNAs,预测和探究养心通脉方有效部位方(active principle region of Yangxing Tongmai formula,apr-YTF)诱导骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)心肌样分化的分子机制。方法:提取急性心肌梗死(acute myocardial infarction,AMI)血瘀证大鼠BMSCs并培养,使用apr-YTF含药血清诱导,以阴性对照血清做对照,提取细胞总RNAs,使用表达谱芯片检测细胞miRNAs的表达,并进行聚类分析。运用生物信息学方法预测差异miRNA调控的靶基因和功能分析。结果:芯片探测出共有26个差异miRNAs,其中8个上调,18个下调,经筛选和预测共100个靶基因参与了19种生物过程,11种细胞成分,10种分子作用。轴突导向通路,癌症转录误调节通路,AMPK信号通路被富集。rno-miR-93-5p,rno-miR-204-5p,rno-miR-128-3p为网络调控中心,其中rno-miR-204-5p下调更显著。结论:养心通脉有效部位方干预大鼠骨髓间充质干细胞心肌样分化的机制可能与rno-miR-204-5p,rno-miR-93-5p,轴突导向通路相关性较高。
OBJECTIVE: To predict and explore the cardiomyocyte-like differentiation of bone marrow mesenchymal stem cells (BMSCs) induced by active principle region of Yangxing Tongmai formula (apr-YTF) by screening differentially expressed miRNAs Molecular mechanism. Methods: BMSCs of acute myocardial infarction (AMI) rats with blood stasis were extracted and cultured, induced by serum containing apr-YTF, and negative control serum was used as control to extract total RNAs of cells. The expression of microRNAs Expression, and cluster analysis. Prediction of Differentially Expressed miRNAs Using Bioinformatics Methods Target Genes and Functional Analysis. Results: A total of 26 differentially expressed miRNAs were detected, of which 8 were up-regulated and 18 were down-regulated. A total of 100 target genes were screened and predicted to participate in 19 biological processes, 11 cell components and 10 molecular effects. Axon guidance pathway, cancer transcription misregulation pathway, AMPK signaling pathway is enriched. rno-miR-93-5p, rno-miR-204-5p and rno-miR-128-3p are the network regulatory centers, of which rno-miR-204-5p down-regulated more significantly. CONCLUSION: The mechanism of Yangxintongmai intervention on rat cardiomyocyte-like differentiation of rat bone marrow mesenchymal stem cells may be related to rno-miR-204-5p, rno-miR-93-5p and axon guidance pathways.