title: Pharmacological modulation of the bradykinin-induced differentiation of human lung fibroblasts: effects of budesonide and formoterol.
abstract: OBJECTIVE
  Bradykinin (BK) induces differentiation of lung fibroblasts into myofibroblasts, which play an important role in extracellular matrix remodeling in the airways of asthmatic patients. It is unclear whether this process is affected by antiasthma therapies. Here, we evaluated whether a glucocorticoid, budesonide (BUD), and a long-acting β2-agonist, formoterol (FM), either alone or in combination, modified BK-induced lung fibroblast differentiation, and affected the BK-activated intracellular signaling pathways.
  
  
  METHODS
  Human fetal lung fibroblasts were incubated with BUD (0.001-0.1 μM) and/or FM (0.0001-0.1 μM) before exposure to BK (0.1 or 1 μM). Fibroblast differentiation into α-smooth-muscle-actin-positive (α-SMA⁺) myofibroblasts, BK2 receptor (B2R) expression, extracellular signal-regulated kinase 1/2 (ERK 1/2) phosphorylation (p-ERK1/2), intracellular Ca²⁺ concentration ([Ca²⁺]i), and p65 nuclear factor kappa B translocation were evaluated.
  
  
  RESULTS
  BUD (0.1 μM) and FM (0.1 μM), either alone or in combination, completely inhibited BK-induced α-SMA protein expression and decreased the numbers of α-SMA⁺ fibroblasts, with a clear reduction in α-SMA stress fibers organization. BUD also completely inhibited the increase of B2R, whereas FM with or without BUD had no effect. BK-induced increases of [Ca²⁺]i and p-ERK1/2 were significantly reduced to similar levels by BUD and FM, either alone or in combination, whereas p65 translocation was completely inhibited by all treatments.
  
  
  CONCLUSIONS
  Both BUD and FM, either alone or in combination, effectively inhibited the BK-induced differentiation of fibroblasts into α-SMA⁺ myofibroblasts and the intracellular signaling pathways involved in fibroblast activation. These results suggest that BUD and FM combination therapy has potential to inhibit fibroblast-dependent matrix remodeling in the airways of asthmatic patients.
authors.affiliation: Pulmonary Disease Unit, G. Gaslini Institute, Genoa, Italy.
authors.forename: Federica
authors.initials: F
authors.lastname: Sabatini
authors.forename: Loredana
authors.initials: L
authors.lastname: Petecchia
authors.forename: Cesare
authors.initials: C
authors.lastname: Usai
authors.forename: Michela
authors.initials: M
authors.lastname: Silvestri
authors.forename: Giovanni A
authors.initials: GA
authors.lastname: Rossi
authors.forename: Anna
authors.initials: A
authors.lastname: Miller-Larsson
authors.forename: Fabio L M
authors.initials: FL
authors.lastname: Ricciardolo
mesh_terms: D001920:Bradykinin; D001993:Bronchodilator Agents / Q000008:administration & dosage / Q000494:pharmacology*; D019819:Budesonide / Q000008:administration & dosage / Q000494:pharmacology*; D002118:Calcium / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D056970:Co-Repressor Proteins / Q000378:metabolism; D004359:Drug Therapy, Combination; D004983:Ethanolamines / Q000008:administration & dosage / Q000494:pharmacology*; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D005347:Fibroblasts / Q000187:drug effects* / Q000378:metabolism; D000068759:Formoterol Fumarate; D006801:Humans; D008168:Lung / Q000166:cytology / Q000187:drug effects*; D058628:Myofibroblasts / Q000378:metabolism; D009687:Nuclear Proteins / Q000378:metabolism; D043782:Receptor, Bradykinin B2 / Q000096:biosynthesis; D015398:Signal Transduction / Q000187:drug effects
chemical_list: D001993:Bronchodilator Agents; D056970:Co-Repressor Proteins; D004983:Ethanolamines; C569368:NKAPL protein, human; D009687:Nuclear Proteins; D043782:Receptor, Bradykinin B2; D019819:Budesonide; D048049:Extracellular Signal-Regulated MAP Kinases; D001920:Bradykinin; D002118:Calcium; D000068759:Formoterol Fumarate