Expression Of MiR-199a-3p In Human Adipocytes Is Regulated By Adipokines And Free Fatty Acids

Objective(1)Investigation the expression of miR-199a-3p in the process of human adipocytes differentiation and in visceral adipose depots from obese subjects.(2)Prediction target genes and functions of miR-199a-3p by bioinformatic analysis.(3)To explore the expression alterations of miR-199a-3p in matured human adipocytes after intervened by adipokines such as leptin,resistin,TNF-a,IL-6 and FFAs.Methods(1)The expression of miR-199a-3p in the human preadipocytes that had differentiated into mature adipocytes was analyzed using quantitative real time PCR.(2)The sequence of miR-199a-3p was acquired from miRBase,NCBI Mapviewer,UCSC Genome Browser database.The target genes of miR-199a-3p were predicted by miRanda、DIANAmT、miRDB、Pictar、Targetscan、miRWalk,and the intersection of the results of five databases with known target gene were further analyzed by GO(gene ontology)and pathway enrichment.Pubmed and google were used to review previous studies of miR-199a-3p.(3)The expression of miR-199a-3p in mature.adipocytes treated with leptin,resistin,TNF-a,IL-6 and FFAs was analyzed using quantitative real time PCR.Results(1)miR-199a-3p expression was significantly upregulated in mature human adipocytes compared to preadipocytes(P<0.05).Additionally,miR-199a-3p expression was higher in visceral adipose depots from obese subjects.(2)miR-199a-3p had some conserved property among different species.By gene ontology analysis,the target genes were enriched in phosphate metabolic process,secretion by cell,response to glucose stimulus,response to monosaccharide stimulus,response to hexose stimulus,response to carbohydrate stimulus,regulation of cell migration,intracellular signaling cascade,protein amino acid dephosphorylation,protein amino acid phosphorylation(P<0.05).(3)By pathway analysis,the target genes were especially involved in regulation of Endocytosis,p53 signaling pathway,ECM-receptor interaction,TGF-beta signaling pathway,Hedgehog signaling pathway,Adherens junction(P<0.05).(4)leptin,TNF-a,IL-6 and FFA significantly promoted miR-199a-3p expression in mature human adipocytes,while resistin had the opposite effect.Conclusions(1)miR-199a-3p may exert a role in modulation of adipocyte differentiation,which may improve adipocyte differentiation.(2)Bioinformatic Analysis results demonstrate miR-199a-3p play a role in adipocytel differentiation,inflammatory reaction and insulin resistance.(3)miR-199a-3p may invovle in the regulation of obesity-related insulin resistance and inflammatory reaction mediated by certain adipokines.