Screening And Bioinformatic Analyses Of Micro Rnas Associated With Adiponectin From Epicardial Adipose Tissue In Coronary Artery Disease

Objective To investigate the differential expression of microRNAs(DE-miRNAs)in patients with coronary atherosclerotic heart disease(CAD)and non-CAD.The differential mi RNAs were analyzed by bioinformatics analysis to find which signal pathway,gene and protein may be involved in these miRNAs regulated adiponectin gene(ADIPOQ)expression.Methods First,the DE-miRNAs of epicardial adipose tissue and plasma of CAD patients or non-CAD patients were screened by microarray technology.Second,to find some target miRNAs which have complementary binding site to match with ADIPOQ 3 '-untranslated region(UTR)or closely relate to ADIPOQ.Then to verificate it by the method of qRT-PCR.Third,the most meaningful two miRNAs(one up-regulation and one down-regulation)were selected for bioinformatics analysis firstly,the target genes of miRNAs were predicted,and then the GO functional enrichment and the KEGG pathway was analyzed by DAVID software,and finally the target protein was introduced into the STRING database to draw the protein-protein interaction network.Results 29 DE-miRNAs(P <0.05,Fold change>2)was screened out from CAD and non-CAD,10 of which were down regulated and 19 up-regulated.Expression of miR-371b-5p and miR-155-5p were verified by the method of qRT-PCR.The results showed that CAD group compared with non-CAD,miR-371b-5p was up-regulated in EAT in patients(P<0.05),while the expression of miR-155-5p was down regulated(P<0.05).The results of bioinformatics analysis showed that the Gene Ontology(GO)function of complementary target genes of miR-371b-5p and miR-155-5p mainly concentrate transcriptional activator activity,RNA polymerase II core promoter proximal region sequence-specific binding,DNA binding,transcription factor activity,protein binding,chromatin binding and other molecular function;the nucleus,nucleoplasm,transcription factor complexes,cytosol and other cell component;regulation of transcription from RNA polymerase II promoter,DNA-templated transcription,protein phosphorylation and other biological processes.The results of KEGG show that the predicted genes of mi R-371b-5p were involved in the 10 pathways,Targeted genes of miR-155-5p are involved in 11 pathways,in which ubiquitin mediated proteolysis,mTOR signaling pathway,TNF pathway,and HIF-1pathway coincide with pathways of miR-371b-5p.ADIPOQ exists in the adipocytokine signaling pathway.STRING protein interaction analysis showed that the predicted gene encoding the protein ADIPOQ,LEP,AKT1,MAPK10,PPARGC1 A,PRKAA1 and SLC2A4 play key roles in maintaining the stability and interaction network,especially ADIPOQ,LEP and AKT1 in the core area of network.Conclusion There are some differential expression miRNAs in EAT or plasma between CAD patients and non-CAD subjects.The expression of miR-371b-5p was up-regulated in EAT tissue and plasma of patients with CAD,while the expression of miR-155-5p was down regulated.The ADIPOQ,LEP and AKT1 might be the downstream targeted genes regulated by miR-371b-5p and miR-155-5p.The network construted by downstream proteins and other related proteins interacting with each other may be involved in regulation of the positive regulation of smooth muscle cell proliferation,adipose tissue development,insulin receptorsignaling pathway and other biological processes.Furthermore,they are involved in the adipocytes signaling pathway,mTOR signal pathway,TNF signaling pathway and HIF-1 signaling pathway,and other signaling pathways.Therefore miR-371b-5p and miR-155-5p play a regulatory role in the formation and development of CAD.