title: Simvastatin Increases the Ability of Roflumilast N-oxide to Inhibit Cigarette Smoke-Induced Epithelial to Mesenchymal Transition in Well-differentiated Human Bronchial Epithelial Cells in vitro.
abstract: BACKGROUND
  Cigarette smoking contributes to epithelial-mesenchymal transition (EMT) in COPD small bronchi as part of the lung remodeling process. We recently observed that roflumilast N-oxide (RNO), the active metabolite of the PDE4 inhibitor roflumilast, prevents cigarette smoke-induced EMT in differentiated human bronchial epithelial cells. Further, statins were shown to protect renal and alveolar epithelial cells from EMT.
  
  
  OBJECTIVE
  To analyze how RNO and simvastatin (SIM) interact on CSE-induced EMT in well-differentiated human bronchial epithelial cells (WD-HBEC) from small bronchi in vitro.
  
  
  METHODS
  WD-HBEC were stimulated with CSE (2.5%). The mesenchymal markers vimentin, collagen type I and α-SMA, the epithelial markers E-cadherin and ZO-1, as well as β-catenin were quantified by real time quantitative PCR or Western blotting. Intracellular reactive oxygen species (ROS) were measured using the H2DCF-DA probe. GTP-Rac1 and pAkt were evaluated by Western blotting.
  
  
  RESULTS
  The combination of RNO at 2 nM and SIM at 100 nM was (over) additive to reverse CSE-induced EMT. CSE-induced EMT was partially mediated by the generation of ROS and the activation of the PI3K/Akt/β-catenin pathway. Both RNO at 2 nM and SIM at 100 nM partially abrogated this pathway, and its combination almost abolished ROS/ PI3K/Akt/β-catenin signaling and therefore EMT.
  
  
  CONCLUSIONS
  The PDE4 inhibitor roflumilast N-oxide acts (over)additively with simvastatin to prevent CSE-induced EMT in WD-HBEC in vitro.
authors.affiliation: 1Clinical Research Unit (UIC), University General Hospital Consortium, Valencia , Spain.
authors.forename: Javier
authors.initials: J
authors.lastname: Milara
authors.forename: Teresa
authors.initials: T
authors.lastname: Peiró
authors.forename: Adela
authors.initials: A
authors.lastname: Serrano
authors.forename: Enrique
authors.initials: E
authors.lastname: Artigues
authors.forename: Jesus
authors.initials: J
authors.lastname: Aparicio
authors.forename: Herman
authors.initials: H
authors.lastname: Tenor
authors.forename: Celia
authors.initials: C
authors.lastname: Sanz
authors.forename: Julio
authors.initials: J
authors.lastname: Cortijo
mesh_terms: D000631:Aminopyridines / Q000494:pharmacology*; D001549:Benzamides / Q000494:pharmacology*; D001980:Bronchi / Q000166:cytology*; D002478:Cells, Cultured; D003521:Cyclopropanes / Q000494:pharmacology; D004847:Epithelial Cells / Q000473:pathology*; D058750:Epithelial-Mesenchymal Transition / Q000187:drug effects*; D006801:Humans; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors / Q000494:pharmacology*; D058988:Phosphodiesterase 4 Inhibitors / Q000494:pharmacology*; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D019821:Simvastatin / Q000494:pharmacology*; D012907:Smoking / Q000009:adverse effects; D051176:beta Catenin / Q000378:metabolism
chemical_list: D000631:Aminopyridines; D001549:Benzamides; D003521:Cyclopropanes; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors; D058988:Phosphodiesterase 4 Inhibitors; D017382:Reactive Oxygen Species; D051176:beta Catenin; D019821:Simvastatin; D051057:Proto-Oncogene Proteins c-akt; C517734:roflumilast N-oxide
keywords: Akt; COPD; cigarette smoke; epithelial mesenchymal transition; phosphatidylinositol 3-kinase; reactive oxygen species; roflumilast N-oxide; simvastatin