Oxidative stress caused by pyocyanin impairs CFTR Cl- transport in human bronchial epithelial cells

Christian Schwarzer, Horst Fischer, Eun Jin Kim, Katharine J. Barber, Aaron D. Mills, Mark J. Kurth, Dieter C. Gruenert, Jung H. Suh, Terry E. Machen, Beate Illek

Research output: Contribution to journalArticle

46 Scopus citations

Abstract

Pyocyanin (N-methyl-1-hydroxyphenazine), a redox-active virulence factor produced by the human pathogen Pseudomonas aeruginosa, is known to compromise mucociliary clearance. Exposure of human bronchial epithelial cells to pyocyanin increased the rate of cellular release of H2O2 threefold above the endogenous H2O2 production. Real-time measurements of the redox potential of the cytosolic compartment using the redox sensor roGFP1 showed that pyocyanin (100 μM) oxidized the cytosol from a resting value of - 318 ± 5 mV by 48.0 ± 4.6 mV within 2 h; a comparable oxidation was induced by 100 μM H2O2. Whereas resting Cl- secretion was slightly activated by pyocyanin (to 10% of maximal currents), forskolin-stimulated Cl- secretion was inhibited by 86%. The decline was linearly related to the cytosolic redox potential (1.8% inhibition/mV oxidation). Cystic fibrosis bronchial epithelial cells homozygous for ΔF508 CFTR failed to secrete Cl- in response to pyocyanin or H2O2, indicating that these oxidants specifically target the CFTR and not other Cl- conductances. Treatment with pyocyanin also decreased total cellular glutathione levels to 62% and cellular ATP levels to 46% after 24 h. We conclude that pyocyanin is a key factor that redox cycles in the cytosol, generates H2O2, depletes glutathione and ATP, and impairs CFTR function in Pseudomonas-infected lungs.

Original languageEnglish (US)
Pages (from-to)1653-1662
Number of pages10
JournalFree Radical Biology and Medicine
Volume45
Issue number12
DOIs
StatePublished - Dec 15 2008

Keywords

  • CFTR
  • Chloride ion transport
  • Cystic fibrosis
  • Free radicals
  • Glutathione
  • Hydrogen peroxide
  • Intracellular redox potential
  • Oxidative stress
  • Pseudomonas aeruginosa
  • Pyocyanin

ASJC Scopus subject areas

  • Biochemistry
  • Physiology (medical)

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    Schwarzer, C., Fischer, H., Kim, E. J., Barber, K. J., Mills, A. D., Kurth, M. J., Gruenert, D. C., Suh, J. H., Machen, T. E., & Illek, B. (2008). Oxidative stress caused by pyocyanin impairs CFTR Cl- transport in human bronchial epithelial cells. Free Radical Biology and Medicine, 45(12), 1653-1662. https://doi.org/10.1016/j.freeradbiomed.2008.09.011