中国药理学通报Issue(7)：940-944,5.DOI:10.3969/j.issn.1001-1978.2015.07.011
缺氧预处理诱导大鼠心肌细胞差异表达microRNA的芯片筛选与生物信息学分析
Screening and bioinformatics analysis of differentially expressed miRNAs induced by hypoxia preconditioning in rat cardiomyocytes
摘要
Abstract
Aim To screen the differentially expressed microRNAs ( miRNAs ) induced by hypoxia precondi-tioning ( HPC ) in adult rat cardiomyocytes, and pre-dict miRNAs-regulated target genes and their func-tions. Methods Cardiomyocytes were isolated from a-dult rat ventricular myocardium and cultured ( in vitro) . The cells were divided into 2 groups: control group ( CON ) and hypoxia preconditioning group ( HPC) . The cardiomyocytes in HPC group were sub-jected to 10 min hypoxia followed by 30 min reoxygen-ation, while the cells in CON group were cultured un-der normal condition. After that, total RNA was ex-tracted and then subjected to miRNA microarray to screen differentially expressed miRNAs. The microar-ray results were further validated by quantitative RT-PCR ( qRT-PCR ) . Bioinformatics analysis was per-formed to predict the miRNAs-regulated target genes and analyze the enriched gene ontology ( GO) and sig-naling pathway ( Pathway) . Results HPC caused sig-nificant changes in miRNAs expression in cardiomyo-cytes as compared to CON group. A total of 12 miR-NAs were up-regulated and 14 miRNAs were down-reg-ulated ( P <0. 01 , FDR <0 . 05 ) . The differentially expressed 7 miRNAs with a fluorescent signal intensity>500 were selected for further bioinformatics analysis. The expression of miR-133b-5p, miR-664-1-5p and miR-6216 detected by qRT-PCR exhibited the similar patterns of up or down regulation to those shown in mi-croarray results. Bioinformatics analysis revealed that miRNAs-regulated target genes were significantly en-riched in 27 GOs and 6 signal pathways. Conclusion <br> The expression profile of miRNAs in rat cardiomyo-cytes is significantly affected by HPC. These differenti-ally expressed miRNAs might participate in HPC-in-duced cardioprotection by regulating their target genes in rat cardiomyocytes.关键词
心肌细胞/成年大鼠/缺氧预处理/microRNA/芯片/生物信息学Key words
cardiomyocytes/adult rat/hypoxia pre-conditioning/microRNA/microarray/bioinformatics分类
医药卫生引用本文复制引用
何淑芳,朱海娟,程洁,许士进,杨婉,张野..缺氧预处理诱导大鼠心肌细胞差异表达microRNA的芯片筛选与生物信息学分析[J].中国药理学通报,2015,(7):940-944,5.基金项目
国家自然科学基金课题( No 81200171) （ No 81200171）
安徽省科技厅年度重点项目( No 1301043030) （ No 1301043030）
高校省级自然科学研究重大项目(No KJ2014ZD16) （No KJ2014ZD16）
