OBJECTIVE: M3 muscarinic acetylcholine receptor (mAChR) plays an important role in the regulation of cytokine production in inflammatory diseases. In this study, we explored the precise role of M3 mAChR under stimulation with agonist in IL-8 expression and of the signaling pathway involved in this process. MATERIALS AND METHODS: Recombinant U2OS cells stably expressing M3 mAChR as a model system were stimulated by carbachol to evaluate the role of M3 mAChR in the expression of IL-8. RESULTS: Activation of M3 mAChR with carbachol increased both IL-8 mRNA and protein expression in a concentration-dependent manner. Elevated IL-8 expression was completely antagonized by atropine, 4-DAMP and tiotropium. M3 mAChR-mediated IL-8 expression was almost completely inhibited by the NF-κB inhibitor BAY11-7082 and, to a lesser extent, by U0126, SB203580, and SP600125, which are inhibitors for ERK1/2, p38, and JNK, respectively. Furthermore, M3 mAChR-mediated NF-κB activation and IL-8 expression were simultaneously attenuated by the PKC inhibitor calphostin C, whereas PMA, a PKC activator, mimicked the effects of carbachol, inducing IL-8 expression. CONCLUSIONS: Our findings offer insights into the specific and critical role of M3 mAChR in regulating inflammatory response and indicate M3 mAChR/PKC/NF-κB signaling axis driven by endogenous acetylcholine as a potential therapeutic targets for inflammatory diseases.
OBJECTIVE: M3 muscarinic acetylcholine receptor (mAChR) plays an important role in the regulation of cytokine production in inflammatory diseases. In this study, we explored the precise role of M3 mAChR under stimulation with agonist in IL-8 expression and of the signaling pathway involved in this process. MATERIALS AND METHODS: Recombinant U2OS cells stably expressing M3 mAChR as a model system were stimulated by carbachol to evaluate the role of M3 mAChR in the expression of IL-8. RESULTS: Activation of M3 mAChR with carbachol increased both IL-8 mRNA and protein expression in a concentration-dependent manner. Elevated IL-8 expression was completely antagonized by atropine, 4-DAMP and tiotropium. M3 mAChR-mediated IL-8 expression was almost completely inhibited by the NF-κB inhibitor BAY11-7082 and, to a lesser extent, by U0126, SB203580, and SP600125, which are inhibitors for ERK1/2, p38, and JNK, respectively. Furthermore, M3 mAChR-mediated NF-κB activation and IL-8 expression were simultaneously attenuated by the PKC inhibitor calphostin C, whereas PMA, a PKC activator, mimicked the effects of carbachol, inducing IL-8 expression. CONCLUSIONS: Our findings offer insights into the specific and critical role of M3 mAChR in regulating inflammatory response and indicate M3 mAChR/PKC/NF-κB signaling axis driven by endogenous acetylcholine as a potential therapeutic targets for inflammatory diseases.
Authors: Silke Appel; Stephanie Le Hellard; Ove Bruland; Johan G Brun; Roald Omdal; Gudlaug Kristjansdottir; Elke Theander; Gunnel Nordmark; Marika Kvarnström; Per Eriksson; Lars Rönnblom; Marie Wahren-Herlenius; Roland Jonsson Journal: Ann Rheum Dis Date: 2011-03-30 Impact factor: 19.103
Authors: Kristen Page; Jing Li; Limei Zhou; Svetlana Iasvovskaia; Kevin C Corbit; Jae-Won Soh; I Bernard Weinstein; Allan R Brasier; Anning Lin; Marc B Hershenson; Svetlana Iasvoyskaia Journal: J Immunol Date: 2003-06-01 Impact factor: 5.422
Authors: Andrew C Kruse; Jianxin Hu; Albert C Pan; Daniel H Arlow; Daniel M Rosenbaum; Erica Rosemond; Hillary F Green; Tong Liu; Pil Seok Chae; Ron O Dror; David E Shaw; William I Weis; Jürgen Wess; Brian K Kobilka Journal: Nature Date: 2012-02-22 Impact factor: 49.962