FFAS And Adipokine-mediated Regulation Of Hsa-miR-143Expression In Human Adipocytes

Objective （1） The different expression of hsa-miR-143in the process ofdifferentiation in human adipocytes was investigated.（2）To predict target gene andfunctions of hsa-miR-143by bioinformatic analysis.（3） The alteration ofhsa-miR-143expression in matured human adipocytes were studied after intervenedby FFAs and some adipokines such as TNF-, IL-6, leptin and resistin.Methods The levels of hsa-miR-143expression in the human preadipocytes thathad differentiated into mature adipocytes were analyzed using quantitative real timePCR. MiRBase, NCBI Mapviewer, UCSC Genome Browser database were used toget the sequence of hsa-miR-143. The target gene were predicted by TargetScan,PicTar and miRanda, and the intersection of results of three database with knowntarget gene were analyzed by Gene Ontology and pathway analysis. Pubmed andgoogle were used to review previous studies of hsa-miR-143. And furthermore, theexpression of hsa-miR-143were measured in mature adipocytes treated with FFAs,TNF-, IL-6, leptin and resistin were analyzed using quantitative real time PCR.Results （1）During the conversion of cultured human preadipocytes to matureadipocytes, the expression of hsa-miR-143was upregulated (P<0.05).（2）Hsa-miR-143was highly conserved among species. By gene ontology analysis, thetarget genes were enriched in actin cytoskeleton organization and biogenesis,positively regulating Rac protein signal transduction, synaptic transmission, and otherbiological processes (P<0.001).（3）By pathway analysis, the target genes weremainly involved in regulation of actin cytoskeleton, Melanoma, Gap junction, MAPKsignaling pathway, and others (P<0.05).（4）The expression of hsa-miR-143were downregulated by FFAs, leptin and resistin in human adipocytes (P<0.05), but not byTNF-and IL-6(P>0.05).Conclusions （1）During the conversion of cultured human preadipocytes to matureadipocytes, the expression of hsa-miR-143is upregulated. Bioinformatic Analysisresults show hsa-miR-143is closely related to cell differentiation and metabolism.（2）hsa-miR-143is inhibited by FFAs,leptin, and resistin,but not by TNF-and IL-6in cultered human adipocytes. These findings indicate that FFAs, leptin, and resistin,may inhibit the expression of miR-143via a negative feedback which requires furtherstudy. In conclusion, we identify the regularity of miR-143expression during theconversion of human preadipocytes into mature adipocytes and several regulativefactors on the miR-143expression in cultured human adipocytes. These data providefurther evidence of the partial involvement of hsa-miR-143in the regulation ofinsulin sensitivity mediated by certain adipokines. Our findings may bring newinsights into the mechanisms of obesity-associated insulin resistance.