Bioinformatics Analysis and Laboratory Study on Innate Immune Signaling Molecule hsa-miR-146b Regulating Stroke-related Signaling Molecules

Abstract: The aim of this study is to predict the stroke-related signaling molecules regulated by innate immunity signaling molecule hsa-miR-146b by the methods of bioinformatics. The UCSC gene browser， TransmiR，miRwalk and Genecards were employed in the present study to figure out the stroke-related signaling molecules regulated by hsa-miR-146b. To verify the miR-146b regulating downstream signaling molecules， use the lentiviral transfection BV2 cells in response to lipopolysaccharide (LPS) and Real-time quantitative PCR to detect of mRNA levels of EGR1， TLR4， miR-146b， BDNF and MMP9. The UCSC gene browser showed that hsa-miR 146b demonstrated high conservatism in several species. Taken all the results together， it was postulated that hsamiR-146b was regulated by the transcription factors EGR1， and at the same time it could regulate the 39 target genes including TLR4， BDNF and MMP9. All the relative genes consisted of a regulatory network in the process of stroke. In LPS stimulated BV2 cells， the mRNA levels of EGR1， TLR4， miR-146b， BDNF and MMP9 were upregulated，while the levels of EGR1， TLR4， miR-146b and MMP9 were decreased in the lentiviral infected BV2 cells，BDNF was increased， indicating that EGR1 was the key signaling molecule regulating miR-146b and downstream signaling molecules as TLR4， BDNF and MMP9. Taken together， the bioinformatics analysis and preliminary experimental verification demonstrated that hsa-miR-146b regulating stroke-related signaling molecules provided prediction for elucidating its role in stroke and valuable guidelines for the further investigation.

CSTR: 32200.14.cjcb.2016.02.0008