P. aeruginosa lipopolysaccharide-induced MUC5AC and CLCA3 expression is partly through Duox1 in vitro and in vivo

PLoS One. 2013 May 14;8(5):e63945. doi: 10.1371/journal.pone.0063945. Print 2013.

Abstract

Background: We have previously found that reactive oxygen species (ROS) are involved in Pseudomonas aeruginosa lipopolysaccharide (PA-LPS) induced MUC5AC in airway epithelial cells. Dual oxidase1 (Duox1), a member of NADPH oxidase(Nox), is known to be responsible for ROS production in respiratory tract epithelial cells. Our aim was to clarify whether Duox1 was also involved in the PA-LPS-induced MUC5AC and calcium dependent chloride channel 3(Clca3), another recognized marker of goblet cell hyperplasia and mucus hyper-production.

Methods: PA-LPS-induced Duox1 mRNA levels were examined in A549 cells, primary mouse tracheal epithelial cells (mTECS) and lung tissues of mice. Nox inhibitors diphenyleneiodonium chloride (DPI) and Duox1 siRNA were used to investigate whether Duox1 is involved in PA-LPS-induced MUC5AC and Clca3 expression both in vitro and in vivo.

Results: Duox1 is induced by PA-LPS in A549 cells, primary mTECs and lung tissues of mice. DPI significantly inhibited PA-LPS-induced up-regulation of Duox1, Muc5ac and Clca3 in primary mouse trachea epithelial cells and lung tissues of mice. Knockdown of Duox1 markedly inhibited PA-LPS-induced MUC5AC expression via a ROS-TGF-α cascade in A549 cells. Furthermore, DPI significantly inhibited PA-LPS-induced increases in inflammatory cells accumulated in mouse lungs.

Conclusions: We demonstrate for the first time that PA-LPS-induced MUC5AC and Clca3 expression is partly through Duox1, and provide supportive evidence for Duox1 as a potential target in treatments of mucin over-production diseases.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Bronchoalveolar Lavage Fluid
  • Cell Line, Tumor
  • Chloride Channels / genetics*
  • Dual Oxidases
  • Epithelial Cells / drug effects
  • Epithelial Cells / metabolism
  • Gene Expression Regulation / drug effects*
  • Gene Knockdown Techniques
  • Humans
  • Lipopolysaccharides / pharmacology*
  • Lung / cytology
  • Mice
  • Mucin 5AC / genetics*
  • Mucoproteins / genetics*
  • NADPH Oxidases / deficiency
  • NADPH Oxidases / genetics*
  • Onium Compounds / pharmacology
  • Pseudomonas aeruginosa / chemistry*
  • RNA, Small Interfering / genetics
  • Reactive Oxygen Species / metabolism
  • Thiourea / analogs & derivatives
  • Thiourea / pharmacology
  • Trachea / cytology
  • Transforming Growth Factor alpha / metabolism

Substances

  • Chloride Channels
  • Clca3a1 protein, mouse
  • Lipopolysaccharides
  • Muc5ac protein, mouse
  • Mucin 5AC
  • Mucoproteins
  • Onium Compounds
  • RNA, Small Interfering
  • Reactive Oxygen Species
  • Transforming Growth Factor alpha
  • diphenyleneiodonium
  • 1,3-dimethylthiourea
  • Dual Oxidases
  • NADPH Oxidases
  • DUOX1 protein, human
  • DUOX2 protein, human
  • Duox1 protein, mouse
  • Thiourea