1. TLR4/NF- κ B-Responsive MicroRNAs and Their Potential Target Genes: A Mouse Model of Skeletal Muscle Ischemia-Reperfusion Injury.
- Author
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Yang, Johnson Chia-Shen, Wu, Shao-Chun, Rau, Cheng-Shyuan, Chen, Yi-Chun, Lu, Tsu-Hsiang, Wu, Yi-Chan, Tzeng, Siou-Ling, Wu, Chia-Jung, and Hsieh, Ching-Hua
- Subjects
CELL receptors ,ALGORITHMS ,ANIMAL experimentation ,BIOLOGICAL models ,CLUSTER analysis (Statistics) ,GENE expression ,GENOMES ,IMMUNOBLOTTING ,MICE ,PROBABILITY theory ,REPERFUSION injury ,RESEARCH funding ,RNA ,SPECTROPHOTOMETRY ,DNA-binding proteins ,BIOINFORMATICS ,OLIGONUCLEOTIDE arrays ,SKELETAL muscle ,GENE expression profiling ,PRECIPITIN tests ,EPIGENOMICS ,CELL physiology - Abstract
Background. The aim of this study was to profile TLR4/NF-κB-responsive microRNAs (miRNAs) and their potential target genes in the skeletal muscles of mice following ischemia-reperfusion injury. Methods. Thigh skeletal muscles of C57BL/6, Tlr4
−/− , and NF-κB−/− mice isolated based on femoral artery perfusion were subjected to ischemia for 2 h and reperfusion for 0 h, 4 h, 1 d, and 7 d. The muscle specimens were analyzed with miRNA arrays. Immunoprecipitation with an argonaute 2- (Ago2-) specific monoclonal antibody followed by whole genome microarray was performed to identify mRNA associated with the RNA-silencing machinery. The potential targets of each upregulated miRNA were identified by combined analysis involving the bioinformatics algorithm miRanda and whole genome expression. Results. Three TLR4/NF-κB-responsive miRNAs (miR-15a, miR-744, and miR-1196) were significantly upregulated in the muscles following ischemia-reperfusion injury. The combined in silico and whole genome microarray approaches identified 5, 4, and 20 potential target genes for miR-15a, miR-744, and miR-1196, respectively. Among the 3 genes (Zbed4, Lrsam1, and Ddx21) regulated by at least 2 of the 3 upregulated miRNAs, Lrsam1 and Ddx21 are known to be associated with the innate immunity pathway. Conclusions. This study profiled TLR4/NF-κB-responsive miRNAs and their potential target genes in mouse skeletal muscle subjected to ischemia-reperfusion injury. [ABSTRACT FROM AUTHOR]- Published
- 2015
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