Thyroid-stimulating hormone stimulates interleukin-6 release from 3T3-L1 adipocytes through a cAMP-protein kinase a pathway

Abstract

Objective: Thyroid-stimulating hormone (TSH) is a novel modulator of adipokine release from human and mouse adipocytes. The aim of our study was to identify the signal transduction pathways activated by TSH that stimulate interleukin (IL)-6 production. Research Methods and Procedures: Mouse 3T3-L1 preadipocyte and differentiated adipocyte cell cultures were studied. The effect of 0 to 1 μM TSH on IL-6 protein release into the medium over 0 to 24 hours was assessed. TSH signaling pathways responsible for regulating IL-6 were studied through the use of 1 μM forskolin, 100 μM 8-pCPT-2′-O-Me- cAMP, 10 μM H89, 50 μM PD98059, and 2 μg/mL actinomycin D. Results: TSH stimulated IL-6 release by 2.6-fold from 3T3-L1 adipocytes at concentrations as low as 0.01 μM but did not alter IL-6 production of corresponding preadipocytes. Forskolin (elevates intracellular cAMP) stimulated IL-6 release from 3T3-L1 adipocytes (n = 3, p < 0.005), and H89, an inhibitor of cAMP-dependent protein kinase A (PKA), reduced TSH-stimulated IL-6 release by 66% (n = 3, p < 0.01), indicating a requirement for cAMP-dependent PKA. Inhibition of the mitogen-activated protein kinase pathway with PD98059 did not affect TSH-stimulated IL-6 release. Activation of an alternate cAMP target, the exchange protein of cAMP, with 8-pCPT-2′-O-Me-cAMP, had no effect on IL-6 release. TSH raised the level of IL-6 mRNA, and blockade of transcription with actinomycin D abrogated IL-6 protein release by TSH (n = 3, p < 0.05). Discussion: TSH stimulates IL-6 release from differentiated 3T3-L1 adipocytes, but not preadipocytes, by signaling through cAMP-PKA to activate IL-6 gene transcription. Copyright © 2006 NAASO.