title: Roflumilast improves corticosteroid resistance COPD bronchial epithelial cells stimulated with toll like receptor 3 agonist.
abstract: BACKGROUND
  Chronic obstructive pulmonary disease (COPD) is characterised by chronic pulmonary inflammation punctuated by periods of viral exacerbations. Recent evidence suggests that the combination of roflumilast with corticosteroids may improve the compromised anti-inflammatory properties of corticosteroids in COPD. We analyzed differential and combination anti-inflammatory effects of dexamethasone and roflumilast N-oxide in human bronchial epithelial cells (HBECs) stimulated with viral toll like receptor (TLR) agonists.
  
  
  METHODS
  Lung tissue and HBECs were isolated from healthy (n = 15), smokers (n = 12) and smokers with COPD (15). TLR3 expression was measured in lung tissue and in HBECs. IL-8 secretion was measured in cell cultures after TLR3 stimulation with poly I:C 10 μg/mL.
  
  
  RESULTS
  We found that TLR3 expression was increased by 1.95 fold (protein) and 2.5 fold (mRNA) in lung tissues from smokers with COPD and inversely correlated with lung function. The TLR3 agonist poly I:C 10 μg/mL increased the IL-8 release in HBECs that was poorly inhibited by dexamethasone in smokers (24.5%) and smokers with COPD (21.6%). In contrast, roflumilast showed similar inhibitory effects on IL-8 release in healthy (58.8%), smokers (56.6%) and smokers with COPD (50.5%). The combination of roflumilast N-oxide and dexamethasone showed additive inhibitory effects. Mechanistically, roflumilast N-oxide when combined with dexamethasone increased the expression of MKP1, and enhanced the inhibitory effects on phospho-p38, AP1 and NFκB activities which may explain the additive anti-inflammatory effects.
  
  
  CONCLUSIONS
  Altogether, our data provide in vitro evidence for a possible clinical utility to add roflumilast on top of inhaled corticosteroid in COPD.
authors: Clinical Research Unit, University General Hospital Consortium, Valencia, Spain. xmilara@hotmail.com.|CIBERES, Health Institute Carlos III, Valencia, Spain. xmilara@hotmail.com.|Pharmacy Department, Fundación de Investigación, University General Hospital Consortium, Avenida tres cruces s/n, Valencia, E-46014, Spain. xmilara@hotmail.com.
authors: Javier
authors: J
authors: Milara
authors: Department of Pharmacology, Faculty of Medicine, University of Valencia, Valencia, Spain. anselm.mg@gmail.com.|Research Foundation, University General Hospital Consortium, Valencia, Spain. anselm.mg@gmail.com.
authors: Anselm
authors: A
authors: Morell
authors: Department of Pharmacology, Faculty of Medicine, University of Valencia, Valencia, Spain. beaballl@etsia.upv.es.
authors: Bea
authors: B
authors: Ballester
authors: Faculty of Biomedic Sciences, European University of Madrid; affiliated center of Valencia, Valencia, Spain. xmilara@gmail.com.
authors: Celia
authors: C
authors: Sanz
authors: Forest Research Institute, Jersey City, NJ, USA. josefreirem@yahoo.com.
authors: Jose
authors: J
authors: Freire
authors: Forest Research Institute, Jersey City, NJ, USA. Max.Qian@frx.com.
authors: Xiaozhong
authors: X
authors: Qian
authors: Forest Research Institute, Jersey City, NJ, USA. maggie_alonso@live.com.
authors: Maggie
authors: M
authors: Alonso-Garcia
authors: Department of Pharmacology, Faculty of Medicine, University of Valencia, Valencia, Spain. Esteban.Morcillo@uv.es.
authors: Esteban
authors: E
authors: Morcillo
authors: Clinical Research Unit, University General Hospital Consortium, Valencia, Spain. julio.cortijo@uv.es.|CIBERES, Health Institute Carlos III, Valencia, Spain. julio.cortijo@uv.es.|Department of Pharmacology, Faculty of Medicine, University of Valencia, Valencia, Spain. julio.cortijo@uv.es.
authors: Julio
authors: J
authors: Cortijo
mesh_terms: D000305:Adrenal Cortex Hormones / Q000494:pharmacology*; D000368:Aged; D000631:Aminopyridines / Q000494:pharmacology*; D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D001549:Benzamides / Q000494:pharmacology*; D001980:Bronchi / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D016022:Case-Control Studies; D002478:Cells, Cultured; D003521:Cyclopropanes / Q000494:pharmacology; D003907:Dexamethasone / Q000494:pharmacology*; D004351:Drug Resistance; D004359:Drug Therapy, Combination; D004847:Epithelial Cells / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D005260:Female; D006801:Humans; D016209:Interleukin-8 / Q000276:immunology / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D011070:Poly I-C / Q000494:pharmacology*; D029424:Pulmonary Disease, Chronic Obstructive / Q000188:drug therapy* / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D012907:Smoking / Q000009:adverse effects; D051196:Toll-Like Receptor 3 / Q000819:agonists* / Q000235:genetics / Q000276:immunology / Q000378:metabolism
chemical_list: D000305:Adrenal Cortex Hormones; D000631:Aminopyridines; D000893:Anti-Inflammatory Agents; D001549:Benzamides; C508606:CXCL8 protein, human; D003521:Cyclopropanes; D016209:Interleukin-8; C495344:TLR3 protein, human; D051196:Toll-Like Receptor 3; D003907:Dexamethasone; C517734:roflumilast N-oxide; D011070:Poly I-C
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