, 2003 and Parris et al., 1999), as well as causing

the dysfunction of pre-synaptic muscarinic (M2) receptors, which enhances the release of acetylcholine (Nie et al., 2009). Chronic exposure to TNF has also been associated with the desensitisation of G-protein coupled receptors (Guo et al., 2005, Kang et al., 2006 and Osawa et al., 2007). The latter two mechanisms have been implicated in asthma pathogenesis. It is noteworthy that other mechanisms may also be involved, as the elevated secretion of TNF causes airway smooth cell contraction by activating different intracellular pathways, depending on pre- and post-transcriptional activity (Tirumurugaan et al., 2007 and Jude et al., 2011). In selleck products addition, we herein show that the TNF action is not dependent on the enhanced protein expression of TNFR1 or TNFR2, which suggests that the elevated concentration of this cytokine in response to in vivo HQ exposure may alter the ability of TNFRs to activate muscarinic receptors, the sensitivity or expression of muscarinic receptors, or subsequent signalling pathways. Mast cell degranulation is a hallmark of airway hyperresponsiveness. Selleckchem RO4929097 Existing data on the mechanism by which TNF promotes mast cell degranulation and consequently, the release of a wide range of smooth muscle cell active mediators, including histamine, cytokines and leukotrienes, is controversial (Brzezińska-Blaszczyk et al., 2000,

Brzezińska-Blaszczyk et al., 2007 and Brzezińska-Blaszczyk and Pietrzak, 1997). Here we show that in vivo HQ exposure

causes CTMC and MMC degranulation that is dependent Astemizole on TNF release, as the pharmacological inhibition of TNF synthesis reduced mast cell degranulation. Furthermore, TNF-induced mast cell degranulation of the HQ-induced tracheal hyperresponsiveness to MCh was further highlighted by the fact that pre-treatment with mast cell stabilizer partially reversed tracheal hyperresponsiveness. The data shown herein strongly suggest that the release of TNF by tracheal epithelium after low levels of HQ exposure triggers airway hyperresponsiveness in response to cholinergic stimulation. In addition, secreted TNF plays an important role in mast cell degranulation, with the subsequent release of chemical mediators that contribute to the maintenance of HQ-induced tracheal hyperresponsiveness. Together, the activation of these pathways may contribute to the development of airway diseases in subjects chronically exposed to HQ, such as smokers and inhabitants of polluted areas. The authors declare that there are no conflicts of interest. The authors thank Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) for financial support (grants no. 08/55382-7; 09/03964-5). Sandra H. P. Farsky and Wothan Tavares de Lima are fellows of the Conselho Nacional de Pesquisa e Tecnologia (CNPq). Simone M.