Inhibition of Mitochondrial Respiration During Early Stage Sepsis

  • Nathan A. Davies
  • Chris E. Cooper
  • Ray Stidwill
  • Mervyn Singer
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 530)


It is known that nitric oxide (NO) is produced in response to a septic insult such as bacterial invasion and that overproduction of NO can have serious debilitating consequences. The mechanism by which NO causes damage at the cellular level is less clear. We have therefore studied the response to a septic insult in an anaesthetised spontaneously breathing Sprague-Dawley rat model. Six rats were given either an intravenous infusion of bacterial cell wall lipopolysaccharide (LPS, 5mg/kg) or saline control over 1 hour. For electron paramagnetic resonance (EPR) studies, blood samples were collected every hour for a further two hours and liver tissue samples were collected postmortem. Measurement was also made of PaO2, blood pressure, base deficit, aortic and renal blood flow and hepatic microvascular pO2 (using porphyrin phosphoresence). Tissue samples were also collected for mitochondrial complex activity analysis. After the administration of LPS blood pressure, blood flow and microvascular PO2 were diminished and the base deficit increased. In addition a clear difference was observed by EPR between control and insulted blood and tissue samples. A large heam-nitrosyl signal is observed as well as an increase in the signal at g=1.94, corresponding to the iron-sulphur centres of complex I becoming more reduced. However no significant difference was observed for any of the mitochondrial complex activities. The effect of the NO produced was to depress the circulatory variables and increase base deficit, combined with a reduced oxygen consumption this implies an impairment of normal aerobic respiration. This was supported by increased iron-sulphur signals observed by EPR indicating a blockage in the mitochondrial redox chain with the subsequent accumulation of electrons. As no effect was observed in the mitochondrial complex activities this indicates that this inhibition is reversible in early stage sepsis. We conclude that nitric oxide produced in response to a septic insult can inhibit mitochondria causing an impairment of oxygen utilisation by aerobic respiration.

Key Words

Nitric oxide sepsis mitochondria electron paramagnetic resonance spectroscopy oxygen utilisation. 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Singer M: Management of multiple organ failure: guidelines but no hard-and-fast rules. J. Antimicrob Chemoth 1998;41:103–112.CrossRefGoogle Scholar
  2. 2.
    Moncada S, Palmer RMJ, Higgs EA: Nitric Oxide: Physiology, Pathophysiology, and Pharmacology. Pharmacol Rev 1991;43:109–142.PubMedGoogle Scholar
  3. 3.
    Kim YM, de Vera ME, Watkins SC, Billiar TR: Nitric oxide protects cultured rat hepatocytes from TNF a-induced apoptosis by inducing heat shock protein 70 expression. J Biol Chem 1997;272:1402–1411.PubMedCrossRefGoogle Scholar
  4. 4.
    Kronke KD, Fehsel K, Kolb-Bachofen V: Nitric oxide: Cytotoxicity versus cytoprotection- How, Why, When, and Where? Nitric Oxide 1997;1:107–120.CrossRefGoogle Scholar
  5. 5.
    Moilanen E, MoilanenT, Knowles R, Charles I, Kadoya Y, AlSaffar N, Revell PA, Moncada S.: Nitric oxide synthase is expressed in human macrophages during foreign body inflammation. Amer J Path 1997;150:881–887.PubMedGoogle Scholar
  6. 6.
    Knowles RG, Merrett M, Salter M, Moncada S: Differential Induction of Brain, Lung and Liver Nitric-Oxide Synthase By Endotoxin in the Rat. Biochem J 1990; 270:833–836.PubMedGoogle Scholar
  7. 7.
    Cipolle MD: Secondary organ dysfunction. From clinical perspectives to molecular mediators. Crit Care Med 1993;9:261–298.Google Scholar
  8. 8.
    Koppenol WH, Moreno JJ, Pryor WA, Ischiropoulos H, Beckman JS: Peroxynitrite, a cloaked oxidant formed by nitric oxide and superoxide. Chem Res Tox 1992;5 834–842.CrossRefGoogle Scholar
  9. 9.
    Cassina A, Radi R: Differential inhibitory action of nitric-oxide and peroxynitrite on mitochiondrial elextron transport. Arch Biochem Biophys 1996;328:309–316.PubMedCrossRefGoogle Scholar
  10. 10.
    Torres J, Darley-Usmar V, Wilson MT: Inhibition of cytochrome c oxidase in turnover by nitric oxide: mechanism and implications for control of respiration. Biochem J 1995; 312:169–173.PubMedGoogle Scholar
  11. 11.
    Poderoso JJ, Carreras MC, Lisdero C, Riobo N, Schopfer F, Boveris A: Nitric-oxide inhibits electrontransfer and increases superoxide radical production in rat-heart mitochondria and submitochondrial particles. Arch Biochem Biophys 1996;328:85–92.PubMedCrossRefGoogle Scholar
  12. 12.
    Rosser DM, Stidwill RP, Jacobson D, Singer M: Oxygen tension in the bladder epithelium rises in both high and low cardiac output endotoxemic spsis. J App Physiol 1995;79:1878–82.Google Scholar
  13. 13.
    Ragan CI, Wilson MT, Darley-Usmar VM, Lowe PN: Sub-fractionation of the mitochondria. In: Darley-Usmar VM, Rickwood D, Wilson MT, eds. Mitochondria, A Practical Approach. Oxford: IRL Press, 1987; 89–105.Google Scholar
  14. 14.
    Peisach J, Blumberg WE, Ogawa S, Rachmilewitz EA, Oltzik R: The Effects of Protein Conformation on the Heme Symmetry in High Spin Ferric Heme Proteins as Studied by Electron Paramagnetic Resonance. J Biol Chem 1971;246:3342–3355.PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2003

Authors and Affiliations

  • Nathan A. Davies
    • 1
  • Chris E. Cooper
    • 1
  • Ray Stidwill
    • 2
  • Mervyn Singer
    • 2
  1. 1.Dept. Biological SciencesUniversity of EssexColchester, EssexUK
  2. 2.Bloomsbury Institute of Intensive Care MedicineUniversity College London Medical SchoolLondonUK

Personalised recommendations