Transcription suppression of peroxisome proliferator-activated receptor gamma2 gene expression by tumor necrosis factor alpha via an inhibition of CCAAT/ enhancer-binding protein delta during the early stage of adipocyte differentiation.

TNFalpha is known to inhibit adipocyte differentiation and induce insulin resistance. Moreover, TNFalpha is known to down-regulate peroxisome proliferator-activated receptor (PPAR)gamma2, an adipocyte-specific nuclear receptor of insulin-sensitizer thiazolidinediones. To clarify molecular mechanisms of TNFalpha- mediated PPARgamma2 down-regulation, we here examined the effect of TNFalpha on transcription regulation of PPARgamma2 gene expression during the early stage of adipocyte differentiation. 3T3-L1 preadipocytes (2 d after 100% confluent) were incubated in a differentiation mixture (dexamethasone, insulin, 3-isobutyl-1-methlxanthine), with or without 50 ng/ml TNFalpha, for 24 h. TNFalpha significantly decreased PPARgamma2 expression both at mRNA and protein levels (to approximately 40%), as well as aP2 mRNA expression. The mouse PPARgamma2 gene promoter region (2.2-kb) was isolated and was used for luciferase reporter assays by transient transfection. TNFalpha significantly suppressed PPARgamma2 gene transcription (to approximately 50%), and deletion analyses demonstrated that the suppression was mediated via CCAAT/enhancer-binding protein (C/EBP) binding elements at the -320/-340 region of the promoter. Moreover, TNFalpha significantly decreased expression of C/EBPdelta mRNA and protein levels (to approximately 40%). EMSA, using 3T3-L1 cells nuclear extracts with the -320/-340 region as a probe, demonstrated the binding of C/EBPdelta to the element, which was significantly decreased by TNFalpha treatment. Overexpression of CEBP/delta prevented the TNFalpha-mediated suppression of PPARgamma2 transactivation. Taken together, TNFalpha suppresses PPARgamma2 gene transcription by the inhibition of C/EBPdelta expression and its DNA binding during the early stage of adipocyte differentiation, which may contribute to the inhibition of adipocyte differentiation, as well as the induction of insulin resistance.