Thyroid-Stimulating Hormone Inhibits Insulin Receptor Substrate-1 Expression and Tyrosyl Phosphorylation in 3T3-L1 Adipocytes by Increasing NF-κB DNA-Binding Activity

Background. Abundant evidence indicates that thyroid-stimulating hormone (TSH) levels are associated with insulin resistance in adipocytes. However, the potential mechanism of the association remains uncertain. The objective of this study was to determine the potential role of TSH in the suppression of insulin receptor substrate-1 (IRS-1) expression and IRS-1 tyrosyl phosphorylation, which might contribute to insulin resistance. Methods. Mouse 3T3-L1 preadipocytes were differentiated into adipocytes. After treatment with 0.01, 0.1, and 1.0 mIU/ml bovine TSH, the TNF-alpha concentration in the medium was determined by enzyme-linked immunosorbent assay (ELISA). Nuclear factor-kappa B (NF-kappa B) DNA-binding activity was quantified by electrophoretic mobility shift assay (EMSA). IRS-1 levels in adipocytes were quantified by Western blotting, and tyrosine phosphorylation was measured by immunoprecipitation. Results. TSH induced TNF-alpha secretion in a dose-dependent manner. There was a significant positive correlation between NF-kappa B DNA-binding activity and TNF-alpha secretion. This effect and correlation were weakened by BAY 11-7082 (a nuclear NF-kappa B inhibitor) and H89 (an inhibitor of cyclic adenosine monophosphate- (cAMP-) dependent protein kinase A (PKA)). Treatment of cultured adipocytes with TSH inhibited insulin-stimulated IRS-1 tyrosyl phosphorylation but promoted TSH-dependent secretion of TNF-alpha and activation of NF-kappa B DNA-binding activity. The effects of TSH were significantly inhibited by BAY 11-7082 and H89 and were completely blocked by the TNF-alpha antagonist WP9QY. Conclusion. TSH inhibited IRS-1 protein expression and tyrosyl phosphorylation in 3T3-L1 adipocytes by stimulating TNF-alpha production via promotion of NF-kappa B DNA-binding activity. TSH might play a pivotal role in the development of insulin resistance.

Automated summary

This study revealed that TSH stimulated the production of TNF-alpha by promoting NF-kappa B DNA-binding activity, inhibiting the expression and tyrosyl phosphorylation of IRS-1 in adipocytes, which may lead to insulin resistance.