Time course of nitric oxide synthases, nitrosative stress, and poly(ADP ribosylation) in an ovine sepsis model

Matthias Lange, Rhykka Connelly, Daniel L. Traber, Atsumori Hamahata, Yoshimitsu Nakano, Aimalohi Esechie, Collette Jonkam, Sanna von Borzyskowski, Lillian D. Traber, Frank C. Schmalstieg, David Herndon, Perenlei Enkhbaatar

Research output: Contribution to journalArticle

38 Citations (Scopus)

Abstract

Introduction: Different isoforms of nitric oxide synthases (NOS) and determinants of oxidative/nitrosative stress play important roles in the pathophysiology of pulmonary dysfunction induced by acute lung injury (ALI) and sepsis. However, the time changes of these pathogenic factors are largely undetermined.Methods: Twenty-four chronically instrumented sheep were subjected to inhalation of 48 breaths of cotton smoke and instillation of live Pseudomonas aeruginosa into both lungs and were euthanized at 4, 8, 12, 18, and 24 hours post-injury. Additional sheep received sham injury and were euthanized after 24 hrs (control). All animals were mechanically ventilated and fluid resuscitated. Lung tissue was obtained at the respective time points for the measurement of neuronal, endothelial, and inducible NOS (nNOS, eNOS, iNOS) mRNA and their protein expression, calcium-dependent and -independent NOS activity, 3-nitrotyrosine (3-NT), and poly(ADP-ribose) (PAR) protein expression.Results: The injury induced severe pulmonary dysfunction as indicated by a progressive decline in oxygenation index and concomitant increase in pulmonary shunt fraction. These changes were associated with an early and transient increase in eNOS and an early and profound increase in iNOS expression, while expression of nNOS remained unchanged. Both 3-NT, a marker of protein nitration, and PAR, an indicator of DNA damage, increased early but only transiently.Conclusions: Identification of the time course of the described pathogenetic factors provides important additional information on the pulmonary response to ALI and sepsis in the ovine model. This information may be crucial for future studies, especially when considering the timing of novel treatment strategies including selective inhibition of NOS isoforms, modulation of peroxynitrite, and PARP.

Original languageEnglish (US)
Article numberR129
JournalCritical Care
Volume14
Issue number4
DOIs
StatePublished - Jul 5 2010

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Nitric Oxide Synthase
Adenosine Diphosphate
Sheep
Sepsis
Lung
Acute Lung Injury
Wounds and Injuries
Protein Isoforms
Poly Adenosine Diphosphate Ribose
Nitric Oxide Synthase Type I
Proteins
Peroxynitrous Acid
Nitric Oxide Synthase Type III
Nitric Oxide Synthase Type II
Smoke
Pseudomonas aeruginosa
Inhalation
DNA Damage
Oxidative Stress
Calcium

ASJC Scopus subject areas

  • Critical Care and Intensive Care Medicine
  • Medicine(all)

Cite this

Time course of nitric oxide synthases, nitrosative stress, and poly(ADP ribosylation) in an ovine sepsis model. / Lange, Matthias; Connelly, Rhykka; Traber, Daniel L.; Hamahata, Atsumori; Nakano, Yoshimitsu; Esechie, Aimalohi; Jonkam, Collette; von Borzyskowski, Sanna; Traber, Lillian D.; Schmalstieg, Frank C.; Herndon, David; Enkhbaatar, Perenlei.

In: Critical Care, Vol. 14, No. 4, R129, 05.07.2010.

Research output: Contribution to journalArticle

Lange, M, Connelly, R, Traber, DL, Hamahata, A, Nakano, Y, Esechie, A, Jonkam, C, von Borzyskowski, S, Traber, LD, Schmalstieg, FC, Herndon, D & Enkhbaatar, P 2010, 'Time course of nitric oxide synthases, nitrosative stress, and poly(ADP ribosylation) in an ovine sepsis model', Critical Care, vol. 14, no. 4, R129. https://doi.org/10.1186/cc9097
Lange, Matthias ; Connelly, Rhykka ; Traber, Daniel L. ; Hamahata, Atsumori ; Nakano, Yoshimitsu ; Esechie, Aimalohi ; Jonkam, Collette ; von Borzyskowski, Sanna ; Traber, Lillian D. ; Schmalstieg, Frank C. ; Herndon, David ; Enkhbaatar, Perenlei. / Time course of nitric oxide synthases, nitrosative stress, and poly(ADP ribosylation) in an ovine sepsis model. In: Critical Care. 2010 ; Vol. 14, No. 4.
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AU - Lange, Matthias

AU - Connelly, Rhykka

AU - Traber, Daniel L.

AU - Hamahata, Atsumori

AU - Nakano, Yoshimitsu

AU - Esechie, Aimalohi

AU - Jonkam, Collette

AU - von Borzyskowski, Sanna

AU - Traber, Lillian D.

AU - Schmalstieg, Frank C.

AU - Herndon, David

AU - Enkhbaatar, Perenlei

PY - 2010/7/5

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N2 - Introduction: Different isoforms of nitric oxide synthases (NOS) and determinants of oxidative/nitrosative stress play important roles in the pathophysiology of pulmonary dysfunction induced by acute lung injury (ALI) and sepsis. However, the time changes of these pathogenic factors are largely undetermined.Methods: Twenty-four chronically instrumented sheep were subjected to inhalation of 48 breaths of cotton smoke and instillation of live Pseudomonas aeruginosa into both lungs and were euthanized at 4, 8, 12, 18, and 24 hours post-injury. Additional sheep received sham injury and were euthanized after 24 hrs (control). All animals were mechanically ventilated and fluid resuscitated. Lung tissue was obtained at the respective time points for the measurement of neuronal, endothelial, and inducible NOS (nNOS, eNOS, iNOS) mRNA and their protein expression, calcium-dependent and -independent NOS activity, 3-nitrotyrosine (3-NT), and poly(ADP-ribose) (PAR) protein expression.Results: The injury induced severe pulmonary dysfunction as indicated by a progressive decline in oxygenation index and concomitant increase in pulmonary shunt fraction. These changes were associated with an early and transient increase in eNOS and an early and profound increase in iNOS expression, while expression of nNOS remained unchanged. Both 3-NT, a marker of protein nitration, and PAR, an indicator of DNA damage, increased early but only transiently.Conclusions: Identification of the time course of the described pathogenetic factors provides important additional information on the pulmonary response to ALI and sepsis in the ovine model. This information may be crucial for future studies, especially when considering the timing of novel treatment strategies including selective inhibition of NOS isoforms, modulation of peroxynitrite, and PARP.

AB - Introduction: Different isoforms of nitric oxide synthases (NOS) and determinants of oxidative/nitrosative stress play important roles in the pathophysiology of pulmonary dysfunction induced by acute lung injury (ALI) and sepsis. However, the time changes of these pathogenic factors are largely undetermined.Methods: Twenty-four chronically instrumented sheep were subjected to inhalation of 48 breaths of cotton smoke and instillation of live Pseudomonas aeruginosa into both lungs and were euthanized at 4, 8, 12, 18, and 24 hours post-injury. Additional sheep received sham injury and were euthanized after 24 hrs (control). All animals were mechanically ventilated and fluid resuscitated. Lung tissue was obtained at the respective time points for the measurement of neuronal, endothelial, and inducible NOS (nNOS, eNOS, iNOS) mRNA and their protein expression, calcium-dependent and -independent NOS activity, 3-nitrotyrosine (3-NT), and poly(ADP-ribose) (PAR) protein expression.Results: The injury induced severe pulmonary dysfunction as indicated by a progressive decline in oxygenation index and concomitant increase in pulmonary shunt fraction. These changes were associated with an early and transient increase in eNOS and an early and profound increase in iNOS expression, while expression of nNOS remained unchanged. Both 3-NT, a marker of protein nitration, and PAR, an indicator of DNA damage, increased early but only transiently.Conclusions: Identification of the time course of the described pathogenetic factors provides important additional information on the pulmonary response to ALI and sepsis in the ovine model. This information may be crucial for future studies, especially when considering the timing of novel treatment strategies including selective inhibition of NOS isoforms, modulation of peroxynitrite, and PARP.

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DO - 10.1186/cc9097

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