title: Doxycycline inhibits TGF-β1-induced extracellular matrix production in nasal polyp-derived fibroblasts.
abstract: BACKGROUND
  Doxycycline has been shown to have antibacterial and anti-inflammatory effects and suppresses collagen biosynthesis. The purpose of this study was to evaluate the effects of doxycycline on transforming growth factor (TGF)-β1-induced myofibroblast differentiation and extracellular matrix production in nasal polyp-derived fibroblasts (NPDFs). We also determined the molecular mechanisms of action for doxycycline.
  
  
  METHODS
  NPDFs were isolated from nasal polyps from 8 patients. Doxycycline was used to pretreat TGF-β1-induced NPDFs. Cytotoxicity was evaluated using a 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyl-tetrazolium bromide assay. Expression levels of α-smooth muscle actin (SMA) and fibronectin were measured using Western blot, reverse-transcription polymerase chain reaction, and immunofluorescence staining. Total collagen production was analyzed with the Sircol collagen assay, while mitogen-activated protein kinase (MAPK) and NF-κB activation were determined using Western blot analysis. Luciferase assay was used to evaluate the transcriptional activity of NF-κB.
  
  
  RESULTS
  Although doxycycline (0 to 40 μg/mL) had no significant cytotoxic effects in TGF-β1-induced NPDFs, it significantly reduced the expression levels of α-SMA, fibronectin, and collagen in TGF-β1-induced NPDFs in a dose-dependent manner. Doxycycline also inhibited the TGF-β1-induced activation of p38, c-Jun NH2 -terminal kinase (JNK), and NF-κB, and its inhibitory effects were similar to those of the specific inhibitors for each.
  
  
  CONCLUSIONS
  Doxycycline has an inhibitory effect on TGF-β1-induced myofibroblast differentiation and extracellular matrix production via the p38 and JNK/NF-κB signal pathways in NPDFs.
authors.affiliation: Department of Otorhinolaryngology-Head and Neck Surgery, Korea University College of Medicine, Seoul, South Korea.
authors.forename: Jae-Min
authors.initials: JM
authors.lastname: Shin
authors.affiliation: Division of Brain Korea 21 Program for Biomedical Science, Korea University College of Medicine, Seoul, South Korea.
authors.forename: Joo-Hoo
authors.initials: JH
authors.lastname: Park
authors.affiliation: Department of Otorhinolaryngology-Head and Neck Surgery, Korea University College of Medicine, Seoul, South Korea.
authors.forename: Il-Ho
authors.initials: IH
authors.lastname: Park
authors.affiliation: Department of Otorhinolaryngology-Head and Neck Surgery, Korea University College of Medicine, Seoul, South Korea.|Division of Brain Korea 21 Program for Biomedical Science, Korea University College of Medicine, Seoul, South Korea.|Research-Driven Hospital, Korea University Guro Hospital, Korea University College of Medicine, Seoul, South Korea.
authors.forename: Heung-Man
authors.initials: HM
authors.lastname: Lee
mesh_terms: D000199:Actins / Q000235:genetics / Q000378:metabolism; D000328:Adult; D000890:Anti-Infective Agents / Q000494:pharmacology*; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D003094:Collagen / Q000235:genetics / Q000378:metabolism; D004318:Doxycycline / Q000494:pharmacology*; D005109:Extracellular Matrix / Q000378:metabolism; D005260:Female; D005347:Fibroblasts / Q000187:drug effects* / Q000502:physiology; D005353:Fibronectins / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D008297:Male; D016328:NF-kappa B / Q000378:metabolism; D009298:Nasal Polyps / Q000188:drug therapy*; D015398:Signal Transduction / Q000187:drug effects; D053773:Transforming Growth Factor beta1 / Q000276:immunology
chemical_list: C541116:ACTA2 protein, human; D000199:Actins; D000890:Anti-Infective Agents; D005353:Fibronectins; D016328:NF-kappa B; D053773:Transforming Growth Factor beta1; D003094:Collagen; D004318:Doxycycline
keywords: MAPK; NF-κB; TGF-β1; doxycycline; extracellular matrix; fibroblast; nasal polyp