Inflammation and airway hyperresponsiveness after chlorine exposure are prolonged by Nrf2 deficiency in mice

Free Radic Biol Med. 2017 Jan:102:1-15. doi: 10.1016/j.freeradbiomed.2016.11.017. Epub 2016 Nov 13.

Abstract

Rationale: Chlorine gas (Cl2) is a potent oxidant and trigger of irritant induced asthma. We explored NF-E2-related factor 2 (Nrf2)-dependent mechanisms in the asthmatic response to Cl2, using Nrf2-deficient mice, buthionine sulfoximine (BSO), an inhibitor of glutathione (GSH) synthesis and sulforaphane (SFN), a phytochemical regulator of Nrf2.

Methods: Airway inflammation and airway hyperresponsiveness (AHR) were assessed 24 and 48h after a 5-min nose-only exposure to 100ppm Cl2 of Nrf2-deficient and wild type Balb/C mice treated with BSO or SFN. Animals were anesthetized, paralyzed and mechanically ventilated (FlexiVent™) and challenged with aerosolized methacholine. Bronchoalveolar lavage (BAL) was performed and lung tissues were harvested for assessment of gene expression.

Results: Cl2 exposure induced a robust AHR and an intense neutrophilic inflammation that, although similar in Nrf2-deficient mice and wild-type mice at 24h after Cl2 exposure, were significantly greater at 48h post exposure in Nrf2-deficient mice. Lung GSH and mRNA for Nrf2-dependent phase II enzymes (NQO-1 and GPX2) were significantly lower in Nrf2-deficient than wild-type mice after Cl2 exposure. BSO reduced GSH levels and promoted Cl2-induced airway inflammation in wild-type mice, but not in Nrf2-deficient mice, whereas SFN suppressed Cl2-induced airway inflammation in wild-type but not in Nrf2-deficient mice. AHR was not affected by either BSO or SFN at 48h post Cl2 exposure.

Conclusions: Nrf2-dependent phase II enzymes play a role in the resolution of airway inflammation and AHR after Cl2 exposure. Moderate deficiency of GSH affects the magnitude of acute inflammation but not AHR.

Keywords: Buthionine sulfoximine; Irritant induced asthma; Nrf2; Phase II enzymes; Sulforaphane.

MeSH terms

  • Animals
  • Bronchoalveolar Lavage
  • Buthionine Sulfoximine / metabolism
  • Chlorine / toxicity
  • Gene Expression Regulation / genetics
  • Glutathione / antagonists & inhibitors
  • Glutathione / biosynthesis
  • Glutathione Peroxidase / genetics
  • Glutathione Peroxidase / metabolism
  • Humans
  • Inflammation / chemically induced
  • Inflammation / metabolism*
  • Inflammation / physiopathology
  • Isothiocyanates / metabolism
  • Lung / drug effects
  • Lung / metabolism*
  • Lung / physiopathology
  • Methacholine Chloride / metabolism
  • Mice
  • NAD(P)H Dehydrogenase (Quinone) / genetics
  • NAD(P)H Dehydrogenase (Quinone) / metabolism
  • NF-E2-Related Factor 2 / genetics*
  • NF-E2-Related Factor 2 / metabolism
  • RNA, Messenger / genetics
  • Respiratory Hypersensitivity / metabolism*
  • Respiratory Hypersensitivity / physiopathology
  • Sulfoxides

Substances

  • Isothiocyanates
  • NF-E2-Related Factor 2
  • Nfe2l2 protein, mouse
  • RNA, Messenger
  • Sulfoxides
  • Methacholine Chloride
  • Chlorine
  • Buthionine Sulfoximine
  • Gpx2 protein, mouse
  • Glutathione Peroxidase
  • NAD(P)H Dehydrogenase (Quinone)
  • Nqo1 protein, mouse
  • sulforaphane
  • Glutathione

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