Screening and bioinformatics analysis of differentially expressed miRNAs induced by hypoxia preconditioning in rat cardiomyocytes
10.3969/j.issn.1001-1978.2015.07.011
- VernacularTitle: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
- Author:
Shufang HE
;
Haijuan ZHU
;
Jie CHENG
;
Shijing XU
;
Wan YAN
;
Ye ZHAGN
- Publication Type:Journal Article
- Keywords:
cardiomyocytes;
adult rat;
hypoxia pre-conditioning;
microRNA;
microarray;
bioinformatics
- From:
Chinese Pharmacological Bulletin
2015;(7):940-944
- CountryChina
- Language:Chinese
-
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
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.
