Metabolic Brain Disease

, Volume 25, Issue 1, pp 23–29 | Cite as

Antioxidant and anti-inflammatory effects of mild hypothermia in the attenuation of liver injury due to azoxymethane toxicity in the mouse

  • Chantal Bémeur
  • Paul Desjardins
  • Roger F. Butterworth
Original Paper


Previous studies have demonstrated protective effects of mild hypothermia following acetaminophen (APAP)-induced acute liver failure (ALF). However, effects of this treatment in ALF due to other toxins have not yet been fully investigated. In the present study, the effects of mild hypothermia in relation to liver pathology, hepatic and cerebral glutathione, plasma ammonia concentrations, progression of encephalopathy, cerebral edema, and plasma proinflammatory cytokines were assessed in mice with ALF resulting from azoxymethane (AOM) hepatotoxicity, a well characterized model of toxic liver injury. Male C57BL/6 mice were treated with AOM (100 µg/g; i.p.) or saline and sacrificed at coma stages of encephalopathy in parallel with AOM mice maintained mildly hypothermic (35°C). AOM treatment led to hepatic damage, significant increase in plasma transaminase activity, decreased hepatic glutathione levels, and brain GSH/GSSG ratios as well as selective increases in expression of plasma proinflammatory cytokines. Mild hypothermia resulted in reduced hepatic damage, improvement in neurological function, normalization of glutathione levels, and selective attenuation in expression of circulating proinflammatory cytokines. These findings demonstrate that the beneficial effects of mild hypothermia in experimental AOM-induced ALF involve both antioxidant and anti-inflammatory mechanisms.


Hepatic encephalopathy Azoxymethane Acute liver failure Hypothermia Brain edema 



Acute liver failure






Blood-brain barrier






Systemic inflammatory response syndrome


Tumor necrosis factor-alpha



This study was supported by a grant from the Canadian Institutes for Health Research. C.B. is a recipient of a fellowship from the Canadian Association for the Study of the Liver (CASL)/Astellas Pharma Canada.


  1. Andus T, Bauer J, Gerok W (1991) Effects of cytokines on the liver. Hepatology 13:364–375CrossRefPubMedGoogle Scholar
  2. Bélanger M, Coté J, Butterworth RF (2006) Neurobiological characterization of an azoxymethane mouse model of acute liver failure. Neurochem Int 48:434–440PubMedGoogle Scholar
  3. Bémeur C, Desjardins P, Qu H, Butterworth RF (2008) N-acetylcysteine selectively modulates inflammatory cytokines in nonacetaminophen acute liver failure in mice. 13th International Symposium on Hepatic Encephalopathy and Nitrogen Metabolism, Padova. Liver Int 28:731, abstractGoogle Scholar
  4. Bémeur C, Desjardins P, Butterworth RF (2010) N-Acetylcysteine attenuates cerebral complications of non-acetaminophen-induced acute liver failure in mice: antioxidant and anti-inflammatory mechanisms. Metab Brain Dis (In press)Google Scholar
  5. Clemmesen JO, Larsen FS, Kondrup J, Hansen BA, Ott P (1999) Cerebral herniation in patients with acute liver failure is correlated with arterial ammonia concentration. Hepatology 29:648–653CrossRefPubMedGoogle Scholar
  6. de Vries HE, Blom-Roosemalen MC, van Oosten M, de Boer AG, van Berkel TJ, Breimer DD, Kuiper J (1996) The influence of cytokines on the integrity of the blood–brain barrier in vitro. J Neuroimmunol 64:37–43CrossRefPubMedGoogle Scholar
  7. Duchini A, Govindarajan S, Santucci M, Zampi G, Hofman FM (1996) Effects of tumor necrosis factor-alpha and interleukin-6 on fluid-phase permeability and ammonia diffusion in CNS-derived endothelial cells. J Investig Med 44:474–482PubMedGoogle Scholar
  8. Ericsson A, Liu C, Hart RP, Sawchenko PE (1995) Type 1 interleukin-1 receptor in the rat brain: distribution, regulation, and relationship to sites of IL-1-induced cellular activation. J Comp Neurol 361:681–698CrossRefPubMedGoogle Scholar
  9. Jalan R, Williams R (2001) The inflammatory basis of intracranial hypertension in acute liver failure. J Hepatol 34:940–942CrossRefPubMedGoogle Scholar
  10. Jalan R, Pollok A, Shah SH, Madhavan K, Simpson KJ (2002) Liver derived proinflammatory cytokines may be important in producing intracranial hypertension in acute liver failure. J Hepatol 37:536–538CrossRefPubMedGoogle Scholar
  11. Jalan R, Olde Damink SW, Deutz NE, Hayes PC, Lee A (2004) Moderate hypothermia in patients with acute liver failure and uncontrolled intracranial hypertension. Gastroenterology 127:1338–1346CrossRefPubMedGoogle Scholar
  12. Jiang W, Desjardins P, Butterworth RF (2009) Direct evidence for central proinflammatory mechanisms in rats with experimental acute liver failure: protective effect of hypothermia. J Cereb Blood Flow Metab 29:944–952CrossRefPubMedGoogle Scholar
  13. Marmarou A, Poll W, Shulman K, Bhagavan H (1978) A simple gravimetric technique for measurement of cerebral edema. J Neurosurg 49:530–537CrossRefPubMedGoogle Scholar
  14. Matkowskyj KA, Marrero JA, Carroll RE, Danilkovich AV, Green RM, Benya RV (1999) Azoxymethane-induced fulminant hepatic failure in C57BL/6J mice: characterization of a new animal model. Am J Physiol 277:G455–G462PubMedGoogle Scholar
  15. Mita A, Hashikura Y, Tagawa Y, Nakayama J, Kawakubo M, Miyagawa S (2005) Expression of Fas ligand by hepatic macrophages in patients with fulminant hepatic failure. Am J Gastroenterol 100:2551–2559CrossRefPubMedGoogle Scholar
  16. Nagaki M, Iwai H, Naiki T, Ohnishi H, Muto Y, Moriwaki H (2000) High levels of serum interleukin-10 and tumor necrosis factor-alpha are associated with fatality in fulminant hepatitis. J Infect Dis 182:1103–1108CrossRefPubMedGoogle Scholar
  17. Riordan SM, Williams R (2003) Mechanisms of hepatocyte injury, multiorgan failure, and prognostic criteria in acute liver failure. Semin Liver Dis 23:203–215CrossRefPubMedGoogle Scholar
  18. Rolando N, Wade J, Davalos M, Wendon J, Philpott-Howard J, Williams R (2000) The systemic inflammatory response syndrome in acute liver failure. Hepatology 32:734–739CrossRefPubMedGoogle Scholar
  19. Rose C, Michalak A, Pannunzio M, Chatauret N, Rambaldi A, Butterworth RF (2000) Mild hypothermia delays the onset of coma and prevents brain edema and extracellular brain glutamate accumulation in rats with acute liver failure. Hepatology 31:872–877CrossRefPubMedGoogle Scholar
  20. Schmidt LE, Larsen FS (2006) Prognostic implications of hyperlactatemia, multiple organ failure, and systemic inflammatory response syndrome in patients with acetaminophen-induced acute liver failure. Crit Care Med 34:337–343CrossRefPubMedGoogle Scholar
  21. Sekiyama KD, Yoshiba M, Thomson AW (1994) Circulating proinflammatory cytokines (IL-1 beta, TNF-alpha, and IL-6) and IL-1 receptor antagonist (IL-1Ra) in fulminant hepatic failure and acute hepatitis. Clin Exp Immunol 98:71–77PubMedCrossRefGoogle Scholar
  22. Simpson KJ (1999) Cytokines, for better or worse? Eur J Gastroenterol Hepatol 11:957–966CrossRefPubMedGoogle Scholar
  23. Tietze F (1969) Enzymic method for quantitative determination of nanogram amounts of total and oxidized glutathione: applications to mammalian blood and other tissues. Anal Biochem 27:502–522CrossRefPubMedGoogle Scholar
  24. Vaquero J, Butterworth RF (2007) Mild hypothermia for the treatment of acute liver failure—what are we waiting for? Nat Clin Pract Gastroenterol Hepatol 4:528–529CrossRefPubMedGoogle Scholar
  25. Vaquero J, Polson J, Chung C, Helenowski I, Schiodt FV, Reisch J, Lee WM, Blei AT (2003) Infection and the progression of hepatic encephalopathy in acute liver failure. Gastroenterology 125:755–764CrossRefPubMedGoogle Scholar
  26. Vaquero J, Rose C, Butterworth RF (2005) Keeping cool in acute liver failure: rationale for the use of mild hypothermia. J Hepatol 43:1067–1077CrossRefPubMedGoogle Scholar
  27. Vaquero J, Bélanger M, James L, Herrero R, Desjardins P, Côté J, Blei AT, Butterworth RF (2007) Mild hypothermia attenuates liver injury and improves survival in mice with acetaminophen toxicity. Gastroenterology 132:372–783CrossRefPubMedGoogle Scholar
  28. Yam LT, Li CY, Crosby WH (1971) Cytochemical identification of monocytes and granulocytes. Am J Clin Pathol 55:283–290PubMedGoogle Scholar
  29. Yasumi Y, Takikawa Y, Endo R, Suzuki K (2007) Interleukin-17 as a new marker of severity of acute hepatic injury. Hepatol Res 37:248–254CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Chantal Bémeur
    • 1
  • Paul Desjardins
    • 1
  • Roger F. Butterworth
    • 1
  1. 1.Neuroscience Research Unit, St-Luc Hospital (CHUM)University of MontrealMontrealCanada

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