Pyocyanin-induced toxicity in A549 respiratory cells is causally linked to oxidative stress

Lee S. Gloyne, Gary D. Grant, Anthony V. Perkins, Catherine M. McDermott, Peter V. Johnson, Gregory J. Anderson, Milton Kiefel, Shailendra Anoopkumar-Dukie, Katie Powell

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Abstract

Pyocyanin, a virulence factor produced by Pseudomonas aeruginosa, has many damaging effects on mammalian cells. Several lines of evidence suggest that this damage is primarily mediated by its ability to generate ROS and deplete host antioxidant defence mechanisms. However, a causal role for oxidative stress has not yet been demonstrated conclusively. Parallel measures of ROS production, antioxidant levels and cytotoxicity provide convincing evidence that pyocyanin-induced cytotoxicity in A549 respiratory cells is mediated by acute ROS production and subsequent oxidative stress. Pyocyanin increased ROS levels in A549 cells as measured by the fluorescent H 2O 2 probes Amplex Red and DCFH-DA. These effects were attenuated by the antioxidant N-acetylcysteine. Furthermore, pyocyanin-induced depletion of intracellular GSH levels 24h after exposure was also prevented by pre-treatment of cells with NAC. Under these conditions, NAC protected cells against pyocyanin-induced cytotoxicity as measured by resazurin reduction to resorufin and viable cell counts, strongly supporting a causal role for oxidative stress. Finally, we also show that pyocyanin-induced activation of the immune and inflammatory transcription factor NF-κB in A549 cells is likely mediated by increased ROS. This increased understanding of mechanisms underlying pyocyanin-induced cytotoxicity may ultimately lead to better strategies for reducing the virulence associated with chronic P. aeruginosa infection.

Original languageEnglish
Pages (from-to)1353-1358
Number of pages6
JournalToxicology in Vitro
Volume25
Issue number7
DOIs
Publication statusPublished - Oct 2011

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Pyocyanine
Oxidative stress
Toxicity
Oxidative Stress
Cytotoxicity
Antioxidants
Pseudomonas aeruginosa
Cells
Pseudomonas Infections
Acetylcysteine
Virulence Factors
A549 Cells
Virulence
Transcription Factors
Cell Count
Chemical activation

Cite this

Gloyne, Lee S. ; Grant, Gary D. ; Perkins, Anthony V. ; McDermott, Catherine M. ; Johnson, Peter V. ; Anderson, Gregory J. ; Kiefel, Milton ; Anoopkumar-Dukie, Shailendra ; Powell, Katie. / Pyocyanin-induced toxicity in A549 respiratory cells is causally linked to oxidative stress. In: Toxicology in Vitro. 2011 ; Vol. 25, No. 7. pp. 1353-1358.
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abstract = "Pyocyanin, a virulence factor produced by Pseudomonas aeruginosa, has many damaging effects on mammalian cells. Several lines of evidence suggest that this damage is primarily mediated by its ability to generate ROS and deplete host antioxidant defence mechanisms. However, a causal role for oxidative stress has not yet been demonstrated conclusively. Parallel measures of ROS production, antioxidant levels and cytotoxicity provide convincing evidence that pyocyanin-induced cytotoxicity in A549 respiratory cells is mediated by acute ROS production and subsequent oxidative stress. Pyocyanin increased ROS levels in A549 cells as measured by the fluorescent H 2O 2 probes Amplex Red and DCFH-DA. These effects were attenuated by the antioxidant N-acetylcysteine. Furthermore, pyocyanin-induced depletion of intracellular GSH levels 24h after exposure was also prevented by pre-treatment of cells with NAC. Under these conditions, NAC protected cells against pyocyanin-induced cytotoxicity as measured by resazurin reduction to resorufin and viable cell counts, strongly supporting a causal role for oxidative stress. Finally, we also show that pyocyanin-induced activation of the immune and inflammatory transcription factor NF-κB in A549 cells is likely mediated by increased ROS. This increased understanding of mechanisms underlying pyocyanin-induced cytotoxicity may ultimately lead to better strategies for reducing the virulence associated with chronic P. aeruginosa infection.",
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Gloyne, LS, Grant, GD, Perkins, AV, McDermott, CM, Johnson, PV, Anderson, GJ, Kiefel, M, Anoopkumar-Dukie, S & Powell, K 2011, 'Pyocyanin-induced toxicity in A549 respiratory cells is causally linked to oxidative stress' Toxicology in Vitro, vol. 25, no. 7, pp. 1353-1358. https://doi.org/10.1016/j.tiv.2011.05.004

Pyocyanin-induced toxicity in A549 respiratory cells is causally linked to oxidative stress. / Gloyne, Lee S.; Grant, Gary D.; Perkins, Anthony V.; McDermott, Catherine M.; Johnson, Peter V.; Anderson, Gregory J.; Kiefel, Milton; Anoopkumar-Dukie, Shailendra; Powell, Katie.

In: Toxicology in Vitro, Vol. 25, No. 7, 10.2011, p. 1353-1358.

Research output: Contribution to journalArticleResearchpeer-review

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