title: β-Adrenergic receptors stimulate interleukin-6 production through Epac-dependent activation of PKCδ/p38 MAPK signalling in neonatal mouse cardiac fibroblasts.
abstract: BACKGROUND AND PURPOSE IL-6 plays crucial roles in cardiac hypertrophy, cardiac fibrosis and heart failure. Activation of β-adrenoceptors induced IL-6 production in neonatal mouse cardiac fibroblasts (NMCFs) through a G(s) /adenylate cyclase/cAMP/p38 MAPK pathway but independent of PKA. However, how cAMP activates p38 MAPK is still not defined. In this study, we have assessed the role of the exchange protein directly activated by cAMP (Epac) and PKCδ in p38 MAPK activation and IL-6 production by stimulated by the β-adrenoceptor agonist isoprenaline in NMCFs. EXPERIMENTAL APPROACH The IL-6 concentration in cell culture supernatants was measured by ELISA. The levels of phosphorylated and total p38 MAPK and PKCδ were determined by Western blot analysis. The translocation of PKCδ was determined by immunoblotting the soluble and particulate fractions. Expression of Epac1 or PKCδ was knocked down by the corresponding, adenovirus-mediated, small hairpin RNA (shRNA). RESULTS In NMCFs, activation of β-adrenoceptors enhanced PKCδ phosphorylation and translocation. Furthermore, knock-down of the PKCδ isoform using an adenovirus-mediated shRNA markedly down-regulated IL-6 induction by NMCFs stimulated with isoprenaline. Moreover, knock-down of Epac1 confirmed that Epac1 was upstream of PKCδ in IL-6 production. Additionally, both Epac1 and PKCδ mediated the p38 MAPK activation induced by isoprenaline. CONCLUSIONS AND IMPLICATIONS β-Adrenoceptor agonists activate a cAMP/Epac/PKCδ/p38 MAPK pathway to produce IL-6 in NMCFs. This study identifies Epac as the link between cAMP and p38 MAPK signalling pathways and demonstrates that PKCδ can function as a novel downstream effector of this β-adrenoceptor/cAMP/Epac pathway.
authors: Institute of Vascular Medicine, Peking University Third Hospital, Key Laboratory of Cardiovascular Molecular Biology and Regulatory Peptides, Ministry of Health and Key Laboratory of Molecular Cardiovascular Science, Ministry of Education, Beijing, China.
authors: Chao
authors: C
authors: Chen
authors: Jianhai
authors: J
authors: Du
authors: Wei
authors: W
authors: Feng
authors: Yao
authors: Y
authors: Song
authors: Zhizhen
authors: Z
authors: Lu
authors: Ming
authors: M
authors: Xu
authors: Zijian
authors: Z
authors: Li
authors: Youyi
authors: Y
authors: Zhang
mesh_terms: D000318:Adrenergic beta-Agonists / Q000494:pharmacology; D000818:Animals; D000831:Animals, Newborn; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000378:metabolism; D005347:Fibroblasts; D020662:Guanine Nucleotide Exchange Factors / Q000378:metabolism*; D015850:Interleukin-6 / Q000378:metabolism*; D007545:Isoproterenol / Q000494:pharmacology; D051379:Mice; D008807:Mice, Inbred BALB C; D009206:Myocardium / Q000166:cytology; D051745:Protein Kinase C-delta / Q000378:metabolism*; D011943:Receptors, Adrenergic, beta / Q000378:metabolism*; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism*
chemical_list: D000318:Adrenergic beta-Agonists; C482601:Epac protein, mouse; D020662:Guanine Nucleotide Exchange Factors; D015850:Interleukin-6; D011943:Receptors, Adrenergic, beta; D000242:Cyclic AMP; D051745:Protein Kinase C-delta; D048051:p38 Mitogen-Activated Protein Kinases; D007545:Isoproterenol