title: Tiotropium bromide inhibits TGF-β-induced MMP production from lung fibroblasts by interfering with Smad and MAPK pathways in vitro.
abstract: BACKGROUND
  chronic obstructive pulmonary disease (COPD) is characterized by chronic inflammation and structural alterations (ie, tissue remodeling) throughout the conducting airways, parenchyma, and pulmonary vasculature. Matrix metalloproteinases (MMPs) are extracellular degrading enzymes that play a critical role in inflammatory cell infiltration and tissue remodeling, but the influence of the agents that are used for the treatment of COPD on the production of MMPs is not well understood.
  
  
  OBJECTIVE
  the present study aimed to examine the influence of tiotropium bromide hydrate (TBH) on the production of MMPs from lung fibroblasts (LFs) induced by transforming growth factor (TGF)-β in vitro.
  
  
  METHODS
  LFs, at a concentration of 5 × 10(5) cells·mL(-1), were stimulated with TGF-β in the presence of various concentrations of TBH. MMP-1 and MMP-2 levels in culture supernatants were examined by enzyme-linked immunosorbent assay (ELISA), and MMP messenger ribonucleic acid (mRNA) expression was examined by real-time polymerase chain reaction (RT-PCR). The influence of TBH on TGF-β signaling pathways was also analyzed by examining Smad activation and signaling protein phosphorylation by ELISA.
  
  
  RESULTS
  TBH at more than 15 pg·mL(-1) inhibited the production of MMP-1 and MMP-2, but not tissue inhibitor of matrix metalloproteinase (TIMP)-1 and TIMP-2, from LFs, after TGF-β stimulation. TBH also suppressed MMP mRNA expression through the inhibition of Smad activation and signaling protein, extracellular-signal-regulated kinase (ERK) 1 and 2, and c-Jun N-terminal kinase (JNK), phosphorylation.
  
  
  CONCLUSIONS
  These results may suggest that TBH suppresses MMP production from LFs, through interference of TGF-β-mediated signaling pathways and results in favorable modification of the clinical status of COPD.
authors.affiliation: Division of Physiology, School of Nursing and Rehabilitation Sciences, Showa University, Midori-ku,Yokohama, Japan. asanok@med.showa-u.ac.jp
authors.forename: Kazuhito
authors.initials: K
authors.lastname: Asano
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Shikama
authors.forename: Naruo
authors.initials: N
authors.lastname: Shoji
authors.forename: Kojiro
authors.initials: K
authors.lastname: Hirano
authors.forename: Harumi
authors.initials: H
authors.lastname: Suzaki
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Nakajima
mesh_terms: D000328:Adult; D000368:Aged; D001483:Base Sequence; D001993:Bronchodilator Agents / Q000037:antagonists & inhibitors* / Q000494:pharmacology*; D018929:Cell Culture Techniques; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005347:Fibroblasts / Q000378:metabolism*; D006801:Humans; D008168:Lung / Q000378:metabolism*; D008297:Male; D020782:Matrix Metalloproteinases / Q000187:drug effects* / Q000378:metabolism; D008875:Middle Aged; D016133:Polymerase Chain Reaction / Q000379:methods; D029424:Pulmonary Disease, Chronic Obstructive; D012602:Scopolamine Derivatives / Q000494:pharmacology*; D051899:Smad2 Protein / Q000187:drug effects* / Q000378:metabolism; D000069447:Tiotropium Bromide; D016212:Transforming Growth Factor beta / Q000187:drug effects* / Q000378:metabolism
chemical_list: D001993:Bronchodilator Agents; D012602:Scopolamine Derivatives; D051899:Smad2 Protein; D016212:Transforming Growth Factor beta; D020782:Matrix Metalloproteinases; D000069447:Tiotropium Bromide