title: Inducible nitric oxide synthase is not required in the development of endotoxin tolerance in mice.
abstract: We investigated the role of inducible nitric oxide synthase (iNOS) in endotoxin tolerance, which was induced in mice genetically deficient of iNOS (iNOS-/-) and in wild-type littermates. In non-tolerant wild-type mice, endotoxin induced high mortality, elevation of plasma levels of nitrite and nitrate, tumor necrosis factor a (TNFalpha), and interleukin 10 (IL-10). These events were preceded by degradation of inhibitors kappaBalpha (IkappaBalpha) and kappaBI (IkappaBbeta), and activation of nuclear factor-kappaB (NF-kappaB) in the lung. Pretreatment of wild-type mice with a sublethal dose of endotoxin prior to lethal endotoxin administration ameliorated lethality and blunted TNFalpha production, whereas IL-10, nitrite, and nitrate production was maintained. These events were associated with reduction of IKBa degradation and NF-kappaB activation in the lung. The kinetics of degradation of IkappaBbeta were also altered. In parallel experiments, nontolerant iNOS-/- mice experienced similar mortality after endotoxin as nontolerant wild-type mice. Plasma levels of nitrite and nitrate were not elevated after lethal endotoxin administration. IL-10 levels were significantly reduced in comparison to nontolerant wild-type mice, whereas TNFalpha levels were similarly increased. These events were preceded by lesser degradation of IkappaBalpha and reduced NF-kappaB activation in the lung. Pretreatment of iNOS-/- mice with a sublethal endotoxin ameliorated lethality. TNFalpha production was significantly reduced, whereas IL-10 production was significantly increased when compared to nontolerant iNOS-/- mice. Degradation of IkappaBalpha and activation of NF-kappaB in the lung were not altered by endotoxin tolerance, whereas kinetics of IkappaBbeta degradation was only delayed. Our data suggests that iNOS is not required for the development of endotoxin tolerance, and that other signal transduction pathways, rather than NF-kappaB, may regulate induction of endotoxin tolerance in the absence of iNOS.
authors.affiliation: Division of Critical Care Medicine, Children's Hospital Medical Center, Cincinnati, Ohio 45229, USA.
authors.forename: Basilia
authors.initials: B
authors.lastname: Zingarelli
authors.forename: Paul W
authors.initials: PW
authors.lastname: Hake
authors.forename: James A
authors.initials: JA
authors.lastname: Cook
mesh_terms: D000818:Animals; D004361:Drug Tolerance / Q000235:genetics / Q000502:physiology; D004731:Endotoxins / Q000633:toxicity*; D005260:Female; D020672:I-kappa B Proteins / Q000378:metabolism; D016753:Interleukin-10 / Q000096:biosynthesis; D008168:Lung / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D000072000:NF-KappaB Inhibitor alpha; D016328:NF-kappa B / Q000378:metabolism; D009566:Nitrates / Q000378:metabolism; D019001:Nitric Oxide Synthase / Q000172:deficiency / Q000235:genetics / Q000502:physiology*; D052247:Nitric Oxide Synthase Type II; D009573:Nitrites / Q000378:metabolism; D012772:Shock, Septic / Q000201:enzymology* / Q000209:etiology / Q000503:physiopathology; D015398:Signal Transduction; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis
chemical_list: D004731:Endotoxins; D020672:I-kappa B Proteins; D016328:NF-kappa B; C000606205:Nfkbia protein, mouse; D009566:Nitrates; D009573:Nitrites; D014409:Tumor Necrosis Factor-alpha; D016753:Interleukin-10; D000072000:NF-KappaB Inhibitor alpha; D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II; C496317:Nos2 protein, mouse