PT  - JOURNAL ARTICLE
AU  - Javier Milara
AU  - Adela Serrano
AU  - Teresa Peirό
AU  - Ricardo Guijarro
AU  - Amadeu Gavaldà
AU  - Montserrat Miralpeix
AU  - Esteban Morcillo
AU  - Julio Cortijo
TI  - Effects of aclidinium on human lung fibroblast activation <em>in vitro</em>
DP  - 2011 Sep 01
TA  - European Respiratory Journal
PG  - 3449
VI  - 38
IP  - Suppl 55
4099  - http://erj.ersjournals.com/content/38/Suppl_55/3449.short
4100  - http://erj.ersjournals.com/content/38/Suppl_55/3449.full
SO  - Eur Respir J2011 Sep 01; 38
AB  - Introduction: Muscarinic activation of human lung fibroblasts is associated with pathological remodelling in the airways of patients with asthma and chronic obstructive pulmonary disease (COPD).Aims: To investigate the in vitro effects of aclidinium bromide, a novel, long-acting muscarinic antagonist, on human lung fibroblast activation.Methods: Lung fibroblasts, isolated from human bronchus, were pre-incubated with aclidinium (10-9M-10-7M), the ERK 1/2 inhibitor PD98059 (10μM) or the cAMP analogue dbcAMP (1mM) for 30 min and then exposed to carbachol (10μM) for 48 h. Collagen type I and α-smooth muscle actin (αSMA) expression were measured by RT-PCR, Western blot (WB) and immunofluorescence. ERK 1/2 phosphorylation was measured by WB and intracellular cAMP levels by cAMP Biotrak enzyme immunoassay. Fibroblast proliferation was assessed using a BrdU kit, and fibroblast migration by wound closure assay.Results: Aclidinium, PD98059 and dbcAMP attenuated carbachol-induced increases in αSMA and collagen type I mRNA and protein levels. Aclidinium and dbcAMP prevented carbachol-induced increases in phospho-ERK 1/2. Carbachol (10μM) prevented isoprenaline (1μM)-induced cAMP upregulation, which was completely reversed by aclidinium 10-7M. Carbachol-dependent increases in lung fibroblast proliferation (2-fold) were reduced by aclidinium 10-7M (1.1-fold), PD98059 (1.3-fold) and dbcAMP (1.2-fold). Aclidinium 10-7M, PD98059 and dbcAMP reduced fibroblast wound closure by 30%, 28% and 40%, respectively.Conclusions: Aclidinium blocks carbachol-induced lung fibroblast proliferation probably by a direct effect at muscarinic receptors. Aclidinium may alleviate lung fibroblast activation in patients with asthma and COPD.