Role of NOS2 in pulmonary injury and repair in response to bleomycin

Changjiang Guo, Elena Atochina-Vasserman, Helen Abramova, Blessy George, Veleeparambil Manoj, Pamela Scott, Andrew Gow

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Nitric oxide (NO) is derived from multiple isoforms of the Nitric Oxide Synthases (NOSs) within the lung for a variety of functions; however, NOS2-derived nitrogen oxides seem to play an important role in inflammatory regulation. In this study, we investigate the role of NOS2 in pulmonary inflammation/fibrosis in response to intratracheal bleomycin instillation (ITB) and to determine if these effects are related to macrophage phenotype. Systemic NOS2 inhibition was achieved by administration of 1400 W, a specific and potent NOS2 inhibitor, via osmotic pump starting six days prior to ITB. 1400 W administration attenuated lung inflammation, decreased chemotactic activity of the broncheoalveolar lavage (BAL), and reduced BAL cell count and nitrogen oxide production. S-nitrosylated SP-D (SNO-SP-D), which has a pro-inflammatory function, was formed in response to ITB; but this formation, as well as structural disruption of SP-D, was inhibited by 1400 W. mRNA levels of IL-1β, CCL2 and Ptgs2 were decreased by 1400 W treatment. In contrast, expression of genes associated with alternate macrophage activation and fibrosis Fizz1, TGF-β and Ym-1 was not changed by 1400 W. Similar to the effects of 1400 W, NOS2-/- mice displayed an attenuated inflammatory response to ITB (day 3 and day 8 post-instillation). The DNA-binding activity of NF-κB was attenuated in NOS2-/- mice; in addition, expression of alternate activation genes (Fizz1, Ym-1, Gal3, Arg1) was increased. This shift towards an increase in alternate activation was confirmed by western blot for Fizz-1 and Gal-3 that show persistent up-regulation 15 days after ITB. In contrast arginase, which is increased in expression at 8 days post ITB in NOS2-/-, resolves by day 15. These data suggest that NOS2, while critical to the development of the acute inflammatory response to injury, is also necessary to control the late phase response to ITB.

Original languageEnglish (US)
Pages (from-to)293-301
Number of pages9
JournalFree Radical Biology and Medicine
Volume91
DOIs
StatePublished - Feb 1 2016

Fingerprint

Bleomycin
Lung Injury
Repair
Pulmonary Surfactant-Associated Protein D
Macrophages
Chemical activation
Therapeutic Irrigation
Pneumonia
Nitric Oxide
Genes
Nitrogen Oxides
Arginase
Macrophage Activation
Pulmonary Fibrosis
Interleukin-1
Nitric Oxide Synthase
Transcriptional Activation
N-((3-(aminomethyl)phenyl)methyl)ethanimidamide
Protein Isoforms
Fibrosis

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Physiology (medical)

Keywords

  • Bleomycin
  • NOS2 inhibitor
  • Pulmonary inflammation
  • S-nitrosylation
  • Surfactant protein-D

Cite this

Guo, Changjiang ; Atochina-Vasserman, Elena ; Abramova, Helen ; George, Blessy ; Manoj, Veleeparambil ; Scott, Pamela ; Gow, Andrew. / Role of NOS2 in pulmonary injury and repair in response to bleomycin. In: Free Radical Biology and Medicine. 2016 ; Vol. 91. pp. 293-301.
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Guo, C, Atochina-Vasserman, E, Abramova, H, George, B, Manoj, V, Scott, P & Gow, A 2016, 'Role of NOS2 in pulmonary injury and repair in response to bleomycin', Free Radical Biology and Medicine, vol. 91, pp. 293-301. https://doi.org/10.1016/j.freeradbiomed.2015.10.417

Role of NOS2 in pulmonary injury and repair in response to bleomycin. / Guo, Changjiang; Atochina-Vasserman, Elena; Abramova, Helen; George, Blessy; Manoj, Veleeparambil; Scott, Pamela; Gow, Andrew.

In: Free Radical Biology and Medicine, Vol. 91, 01.02.2016, p. 293-301.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Role of NOS2 in pulmonary injury and repair in response to bleomycin

AU - Guo, Changjiang

AU - Atochina-Vasserman, Elena

AU - Abramova, Helen

AU - George, Blessy

AU - Manoj, Veleeparambil

AU - Scott, Pamela

AU - Gow, Andrew

PY - 2016/2/1

Y1 - 2016/2/1

N2 - Nitric oxide (NO) is derived from multiple isoforms of the Nitric Oxide Synthases (NOSs) within the lung for a variety of functions; however, NOS2-derived nitrogen oxides seem to play an important role in inflammatory regulation. In this study, we investigate the role of NOS2 in pulmonary inflammation/fibrosis in response to intratracheal bleomycin instillation (ITB) and to determine if these effects are related to macrophage phenotype. Systemic NOS2 inhibition was achieved by administration of 1400 W, a specific and potent NOS2 inhibitor, via osmotic pump starting six days prior to ITB. 1400 W administration attenuated lung inflammation, decreased chemotactic activity of the broncheoalveolar lavage (BAL), and reduced BAL cell count and nitrogen oxide production. S-nitrosylated SP-D (SNO-SP-D), which has a pro-inflammatory function, was formed in response to ITB; but this formation, as well as structural disruption of SP-D, was inhibited by 1400 W. mRNA levels of IL-1β, CCL2 and Ptgs2 were decreased by 1400 W treatment. In contrast, expression of genes associated with alternate macrophage activation and fibrosis Fizz1, TGF-β and Ym-1 was not changed by 1400 W. Similar to the effects of 1400 W, NOS2-/- mice displayed an attenuated inflammatory response to ITB (day 3 and day 8 post-instillation). The DNA-binding activity of NF-κB was attenuated in NOS2-/- mice; in addition, expression of alternate activation genes (Fizz1, Ym-1, Gal3, Arg1) was increased. This shift towards an increase in alternate activation was confirmed by western blot for Fizz-1 and Gal-3 that show persistent up-regulation 15 days after ITB. In contrast arginase, which is increased in expression at 8 days post ITB in NOS2-/-, resolves by day 15. These data suggest that NOS2, while critical to the development of the acute inflammatory response to injury, is also necessary to control the late phase response to ITB.

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Guo C, Atochina-Vasserman E, Abramova H, George B, Manoj V, Scott P et al. Role of NOS2 in pulmonary injury and repair in response to bleomycin. Free Radical Biology and Medicine. 2016 Feb 1;91:293-301. https://doi.org/10.1016/j.freeradbiomed.2015.10.417