Time course of nitric oxide synthases, nitrosative stress, and poly(ADP ribosylation) in an ovine sepsis model.
BACKGROUND
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.
Lange
Matthias
M
Department of Anesthesiology, Investigational Intensive Care Unit, The University of Texas Medical Branch and Shriners Burns Hospital for Children, 301 University Boulevard, Galveston, Texas 77550, USA. lanm@gmx.de
Connelly
Rhykka
R
Traber
Daniel L
DL
Hamahata
Atsumori
A
Nakano
Yoshimitsu
Y
Esechie
Aimalohi
A
Jonkam
Collette
C
von Borzyskowski
Sanna
S
Traber
Lillian D
LD
Schmalstieg
Frank C
FC
Herndon
David N
DN
Enkhbaatar
Perenlei
P
D055371:Acute Lung Injury / Q000378:metabolism / Q000503:physiopathology; D000818:Animals; D004195:Disease Models, Animal; D016209:Interleukin-8 / Q000032:analysis / Q000378:metabolism / Q000502:physiology; D008168:Lung / Q000737:chemistry / Q000201:enzymology; D009566:Nitrates / Q000097:blood; D019001:Nitric Oxide Synthase / Q000378:metabolism* / Q000502:physiology; D052247:Nitric Oxide Synthase Type II / Q000378:metabolism / Q000502:physiology; D052250:Nitric Oxide Synthase Type III / Q000378:metabolism / Q000502:physiology; D009573:Nitrites / Q000097:blood; D011064:Poly Adenosine Diphosphate Ribose / Q000032:analysis / Q000378:metabolism* / Q000502:physiology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D018805:Sepsis / Q000378:metabolism* / Q000503:physiopathology; D012756:Sheep; D013997:Time Factors; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000032:analysis / Q000378:metabolism / Q000502:physiology
D016209:Interleukin-8; D009566:Nitrates; D009573:Nitrites; D011064:Poly Adenosine Diphosphate Ribose; C002744:3-nitrotyrosine; D014443:Tyrosine; D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II; D052250:Nitric Oxide Synthase Type III