Aclidinium inhibits human lung fibroblast to myofibroblast transition

Background Fibroblast to myofibroblast transition is believed to contribute to airway remodelling in lung diseases such as asthma and chronic obstructive pulmonary disease. This study examines the role of aclidinium, a new long-acting muscarinic antagonist, on human fibroblast to myofibroblast transition. Methods Human bronchial fibroblasts were stimulated with carbachol (10(-8) to 10(-5) M) or transforming growth factor-beta 1 (TGF-beta 1; 2 ng/ml) in the presence or absence of aclidinium (10(-9) to 10(-7) M) or different drug modulators for 48 h. Characterisation of myofibroblasts was performed by analysis of collagen type I and a-smooth muscle actin (alpha-SMA) mRNA and protein expression as well as alpha-SMA microfilament immunofluorescence. ERK1/2 phosphorylation, RhoA-GTP and muscarinic receptors (M) 1, 2 and 3 protein expression were determined by western blot analysis and adenosine 3'-5' cyclic monophosphate levels were determined by ELISA. Proliferation and migration of fibroblasts were also assessed. Results Collagen type I and alpha-SMA mRNA and protein expression, as well as percentage alpha-SMA microfilament-positive cells, were upregulated in a similar way by carbachol and TGF-beta 1, and aclidinium reversed these effects. Carbachol-induced myofibroblast transition was mediated by an increase in ERK1/2 phosphorylation, RhoA-GTP activation and cyclic monophosphate downregulation as well as by the autocrine TGF-beta 1 release, which were effectively reduced by aclidinium. TGF-beta 1 activated the non-neuronal cholinergic system. Suppression of M1, M2 or M3 partially prevented carbachol-and TGF-beta 1-induced myofibroblast transition. Aclidinium dose-dependently reduced fibroblast proliferation and migration. Conclusion Aclidinium inhibits human lung fibroblast to myofibrobast transition.