title: Aclidinium inhibits human lung fibroblast to myofibroblast transition.
abstract: 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-β1 (TGF-β1; 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 α-smooth muscle actin (α-SMA) mRNA and protein expression as well as α-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 α-SMA mRNA and protein expression, as well as percentage α-SMA microfilament-positive cells, were upregulated in a similar way by carbachol and TGF-β1, 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-β1 release, which were effectively reduced by aclidinium. TGF-β1 activated the non-neuronal cholinergic system. Suppression of M1, M2 or M3 partially prevented carbachol- and TGF-β1-induced myofibroblast transition. Aclidinium dose-dependently reduced fibroblast proliferation and migration.
  
  
  CONCLUSIONS
  Aclidinium inhibits human lung fibroblast to myofibrobast transition.
authors.affiliation: Unidad de Investigación, Consorcio Hospital General, Universitario, Avenida Tres Vruces s/n, Valencia E-46014, Spain. xmilara@hotmail.com
authors.forename: Javier
authors.initials: J
authors.lastname: Milara
authors.forename: Adela
authors.initials: A
authors.lastname: Serrano
authors.forename: Teresa
authors.initials: T
authors.lastname: Peiró
authors.forename: Amadeu
authors.initials: A
authors.lastname: Gavaldà
authors.forename: Montserrat
authors.initials: M
authors.lastname: Miralpeix
authors.forename: Esteban Jesús
authors.initials: EJ
authors.lastname: Morcillo
authors.forename: Julio
authors.initials: J
authors.lastname: Cortijo
mesh_terms: D000199:Actins / Q000096:biosynthesis / Q000235:genetics; D001980:Bronchi / Q000166:cytology*; D002217:Carbachol / Q000037:antagonists & inhibitors / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D018679:Cholinergic Agonists / Q000494:pharmacology; D024042:Collagen Type I / Q000096:biosynthesis / Q000235:genetics; D004305:Dose-Response Relationship, Drug; D005347:Fibroblasts / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D018727:Muscarinic Antagonists / Q000494:pharmacology*; D058628:Myofibroblasts / Q000187:drug effects*; D012333:RNA, Messenger / Q000235:genetics; D053773:Transforming Growth Factor beta1 / Q000037:antagonists & inhibitors / Q000494:pharmacology; D014326:Tropanes / Q000494:pharmacology*; D015854:Up-Regulation / Q000187:drug effects
chemical_list: D000199:Actins; D018679:Cholinergic Agonists; D024042:Collagen Type I; D018727:Muscarinic Antagonists; D012333:RNA, Messenger; D053773:Transforming Growth Factor beta1; D014326:Tropanes; D002217:Carbachol; C542859:aclidinium bromide