[Prediction of regulating network of innate immune signaling molecule hsa-miR-181a in stroke development based on bioinformatics analysis].
OBJECTIVE
To predict the regulating network of innate immunity signaling molecule hsa-miR-181a in stroke based on the methods of bioinformatics.


METHODS
The UCSC genome browser, the human miRNA disease database (HMDD), the transcription factor-miRNA regulation database (TransmiR), the database on predicted and validated miRNA targets (miRwalk), the Genecards, the long non-coding RNA (LncRNA) disease database, the DIANA LAB-LncBase and the ConSite were employed to study the upstream transcription factor, downstream target genes and the interactive LncRNA of hsa-miR-181a and to draw the core regulating network of hsa-miR-181a. To verify the hsa-miR-181a regulating network, we used lipopolysaccharide (LPS) to stimulate the BV2 cells transfected by lentivirus and real-time quantitative PCR to detect the changes of Toll-like receptor 4 (TLR4), tumor protein 63 (p63), miR-181a and nuclear factor κB (NF-κB) p65.


RESULTS
The UCSC genome browser showed that hsa-miR-181a had two subtypes, which were demonstrated with high conservatism in several species. Diseases analysis and literatures investigation revealed that the hsa-miR-181a was related with many diseases, especially ischemia diseases. Bioinformatics analysis indicated that hsa-miR-181a was regulated by the transcription factors p63, and at the same time, it could regulate 58 target genes such as brain-derived neurotrophic factor (BDNF), TLR4 etc. IncRNA CDKN2B-AS1 and its transcription factors Snail and n-MYC might also interact with hsa-miR-181a. All the relative genes composed a regulatory network with hsa-miR-181a as a core and played important roles in the process of stroke. In LPS-stimulated BV2 cells, the expression levels of TLR4, p63, miR-181a were up-regulated; while the levels of p63, miR-181a and NF-κB p65 decreased in the lentivirus-infected BV2 cells, indicating that p63 was the key signaling molecule in the process of TLR4 regulating miR-181a.


CONCLUSIONS
The bioinformatics analysis and preliminary experimental verification predicted and demonstrated the regulating network of hsa-miR-181a in stroke.
Department of Pharmacy, First Affiliated Hospital, Nanchang University, Nanchang 330046, China, China.
Yanni
Y
Lyu
Research Center for Cardiovascular Diseases, Institute of Translational Medicine, Nanchang University, Nanchang 330031, China.
Yisong
Y
Qian
Department of Pharmacy, First Affiliated Hospital, Nanchang University, Nanchang 330046, China, China.
Longsheng
L
Fu
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002878:Chromosomes, Human, Pair 1 / Q000235:genetics; D019295:Computational Biology / Q000379:methods*; D005786:Gene Expression Regulation / Q000187:drug effects; D053263:Gene Regulatory Networks; D020022:Genetic Predisposition to Disease / Q000235:genetics; D006801:Humans; D007113:Immunity, Innate / Q000235:genetics*; D008070:Lipopolysaccharides / Q000494:pharmacology; D035683:MicroRNAs / Q000235:genetics*; D017628:Microglia / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D008969:Molecular Sequence Data; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012689:Sequence Homology, Nucleic Acid; D015398:Signal Transduction / Q000235:genetics*; D020521:Stroke / Q000235:genetics*; D051197:Toll-Like Receptor 4 / Q000235:genetics; D051996:Transcription Factor RelA / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics; D025521:Tumor Suppressor Proteins / Q000235:genetics
D008070:Lipopolysaccharides; C540193:MIrn181 microRNA, human; D035683:MicroRNAs; C495345:TLR4 protein, human; D051197:Toll-Like Receptor 4; D051996:Transcription Factor RelA; D014157:Transcription Factors; D025521:Tumor Suppressor Proteins