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Circulating acylcarnitines as biomarkers of mitochondrial dysfunction after acetaminophen overdose in mice and humans

  • Molecular Toxicology
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Abstract

Acetaminophen (APAP) is a widely used analgesic. However, APAP overdose is hepatotoxic and is the primary cause of acute liver failure in the developed world. The mechanism of APAP-induced liver injury begins with protein binding and involves mitochondrial dysfunction and oxidative stress. Recent efforts to discover blood biomarkers of mitochondrial damage have identified increased plasma glutamate dehydrogenase activity and mitochondrial DNA concentration in APAP overdose patients. However, a problem with these markers is that they are too large to be released from cells without cell death or loss of membrane integrity. Metabolomic studies are more likely to reveal biomarkers that are useful at early time points, before injury begins. Similar to earlier work, our metabolomic studies revealed that acylcarnitines are elevated in serum from mice after treatment with toxic doses of APAP. Importantly, a comparison with furosemide demonstrated that increased serum acylcarnitines are specific for mitochondrial dysfunction. However, when we measured these compounds in plasma from humans with liver injury after APAP overdose, we could not detect any significant differences from control groups. Further experiments with mice showed that N-acetylcysteine, the antidote for APAP overdose in humans, can reduce acylcarnitine levels in serum. Altogether, our data do not support the clinical measurement of acylcarnitines in blood after APAP overdose due to the standard N-acetylcysteine treatment in patients, but strongly suggest that acylcarnitines would be useful mechanistic biomarkers in other forms of liver injury involving mitochondrial dysfunction.

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Abbreviations

ALT:

Alanine aminotransferase

APAP:

Acetaminophen

CACT:

Carnitine-acylcarnitine translocase

CPT:

Carnitine palmitoyltransferase

GDH:

Glutamate dehydrogenase

JNK:

c-Jun N-terminal kinase

MLK3:

Mixed lineage kinase 3

MPT:

Mitochondrial membrane permeability transition

mtDNA:

Mitochondrial DNA

NAC:

N-acetylcysteine

NAPQI:

N-acetyl-p-benzoquinone imine

OPLS-DA:

Orthogonal projection to latent structures-discriminant analysis

PCA:

Principal component analysis

ROS:

Reactive oxygen species

References

  • Antoine DJ, Dear JW, Starkey-Lewis P, Platt V, Coyle J, Masson M, Thanacoody RH, Gray AJ, Webb DJ, Moggs JG, Bateman DN, Goldring CE, Park BK (2013) Mechanistic biomarkers provide early and sensitive detection of acetaminophen-induced acute liver injury at first presentation to hospital. Hepatology 58:777–787

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Bajt ML, Knight TR, Farhood A, Jaeschke H (2003) Scavenging peroxynitrite with glutathione promotes regeneration and enhances survival during acetaminophen-induced liver injury in mice. J Pharmacol Exp Ther 307:67–73

    Article  CAS  PubMed  Google Scholar 

  • Bajt ML, Knight TR, Lemasters JJ, Jaeschke H (2004) Acetaminophen-induced oxidant stress and cell injury in cultured mouse hepatocytes: protection by N-acetylcysteine. Toxicol Sci 80:343–349

    Article  CAS  PubMed  Google Scholar 

  • Bajt ML, Cover C, Lemasters JJ, Jaeschke H (2006) Nuclear translocation of endonuclease G and apoptosis-inducing factor during acetaminophen-induced liver cell injury. Toxicol Sci 94:217–225

    Article  CAS  PubMed  Google Scholar 

  • Bajt ML, Farhood A, Lemasters JJ, Jaeschke H (2008) Mitochondrial bax translocation accelerates DNA fragmentation and cell necrosis in a murine model of acetaminophen hepatotoxicity. J Pharmacol Exp Ther 324:8–14

    Article  CAS  PubMed  Google Scholar 

  • Bajt ML, Ramachandran A, Yan HM, Lebofsky M, Farhood A, Lemasters JJ, Jaeschke H (2011) Apoptosis-inducing factor modulates mitochondrial oxidant stress in acetaminophen hepatotoxicity. Toxicol Sci 122:598–605

    Article  CAS  PubMed  Google Scholar 

  • Bi H, Li F, Krausz KW, Qu A, Johnson CH, Gonzalez FJ (2013) Targeted metabolomics of serum acylcarnitines evaluates hepatoprotective effect of wuzhi tablet (Schisandra sphenanthera extract) against acute acetaminophen toxicity. Evid Based Complement Alternat Med 2013:985257

    PubMed Central  PubMed  Google Scholar 

  • Chen C, Krausz KW, Shah YM, Idle JR, Gonzalez FJ (2009) Serum metabolomics reveals irreversible inhibition of fatty acid beta-oxidation through the suppression of PPARalpha activation as a contributing mechanism of acetaminophen-induced hepatotoxicity. Chem Res Toxicol 22:699–707

    Article  PubMed Central  PubMed  Google Scholar 

  • Cover C, Mansouri A, Knight TR, Bajt ML, Lemasters JJ, Pessayre D, Jaeschke H (2005) Peroxynitrite-induced mitochondrial and endonuclease-mediated nuclear DNA damage in acetaminophen hepatotoxicity. J Pharmacol Exp Ther 315:879–887

    Article  CAS  PubMed  Google Scholar 

  • Dahlin DC, Miwa GT, Lu AY, Nelson SD (1984) N-acetyl-p-benzoquinone imine: a cytochrome P-450-mediated oxidation product of acetaminophen. Proc Natl Acad Sci USA 81:1327–1331

    Article  CAS  PubMed  Google Scholar 

  • Davern TJ II, James LP, Hinson JA, Polson J, Larson AM, Fontana RJ, Lalani E, Munoz S, Shakil AO, Lee WM, Acute Liver Failure Study Group (2006) Measurement of serum acetaminophen-protein adducts in patients with acute liver failure. Gastroenterology 130:687–694

    Article  CAS  PubMed  Google Scholar 

  • Davis M, Ideo G, Harrison NG, Williams R (1975) Hepatic glutathione depletion and impaired bromosulphthalein clearance early after paracetamol overdose in man and the rat. Clin Sci Mol Med 49:495–502

    CAS  PubMed  Google Scholar 

  • Gunawan BK, Liu ZX, Han D, Hanawa N, Gaarde WA, Kaplowitz N (2006) c-Jun N-terminal kinase plays a major role in murine acetaminophen hepatotoxicity. Gastroenterology 131:165–178

    Article  CAS  PubMed  Google Scholar 

  • Hanawa N, Shinohara M, Saberi B, Gaarde WA, Han D, Kaplowitz N (2008) Role of JNK translocation to mitochondria leading to inhibition of mitochondria bioenergetics in acetaminophen-induced liver injury. J Biol Chem 283:13565–13577

    Article  CAS  PubMed  Google Scholar 

  • Jaeschke H (1990) Glutathione disulfide formation and oxidant stress during acetaminophen-induced hepatotoxicity in mice in vivo: the protective effect of allopurinol. J Pharmacol Exp Ther 255:935–941

    CAS  PubMed  Google Scholar 

  • Jaeschke H, McGill MR (2013) Serum glutamate dehydrogenase—biomarker for liver cell death or mitochondrial dysfunction? Toxicol Sci 134:221–222

    Article  CAS  PubMed  Google Scholar 

  • Jaeschke H, McGill MR, Ramachandran A (2012) Oxidant stress, mitochondria, and cell death mechanisms in drug-induced liver injury: lessons learned from acetaminophen hepatotoxicity. Drug Metab Rev 44:88–106

    Article  CAS  PubMed  Google Scholar 

  • Kaufman DW, Kelly JP, Rosenberg L, Anderson TE, Mitchell AA (2002) Recent patterns of medication use in the ambulatory adult population of the United States: the Slone survey. JAMA 287:337–344

    Article  PubMed  Google Scholar 

  • Knight TR, Ho YS, Farhood A, Jaeschke H (2002) Peroxynitrite is a critical mediator of acetaminophen hepatotoxicity in murine livers: protection by glutathione. J Pharmacol Exp Ther 303:468–475

    Article  CAS  PubMed  Google Scholar 

  • Kon K, Kim JS, Jaeschke H, Lemasters JJ (2004) Mitochondrial permeability transition in acetaminophen-induced necrosis and apoptosis of cultured mouse hepatocytes. Hepatology 40:1170–1179

    Article  CAS  PubMed  Google Scholar 

  • Lauterburg BH, Mitchell JR (1987) Therapeutic doses of acetaminophen stimulate turnover of cysteine and glutathione in man. J Hepatol 4:206–211

    Article  CAS  PubMed  Google Scholar 

  • McGill MR, Jaeschke H (2013) Metabolism and disposition of acetaminophen: recent advances in relation to hepatotoxicity and diagnosis. Pharm Res 30:2174–2187

    Google Scholar 

  • McGill MR, Yan HM, Ramachandran A, Murray GJ, Rollins DE, Jaeschke H (2011) HepaRG cells: a human model to study mechanisms of acetaminophen hepatotoxicity. Hepatology 53:974–982

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • McGill MR, Sharpe MR, Williams CD, Taha M, Curry SC, Jaeschke H (2012) The mechanism underlying acetaminophen-induced hepatotoxicity in humans and mice involves mitochondrial damage and nuclear DNA fragmentation. J Clin Invest 122:1574–1583

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • McGill MR, Lebofsky M, Norris HK, Slawson MH, Bajt ML, Xie Y, Williams CD, Wilkins DG, Rollins DE, Jaeschke H (2013) Plasma and liver acetaminophen-protein adduct levels in mice after acetaminophen treatment: dose-response, mechanisms, and clinical implications. Toxicol Appl Pharmacol 269:240–249

    Article  CAS  PubMed  Google Scholar 

  • Mitchell JR, Jollow DJ, Potter WZ, Gillette JR, Brodie BB (1973) Acetaminophen-induced hepatic necrosis. IV. Protective role of glutathione. J Pharmacol Exp Ther 187:211–217

    CAS  PubMed  Google Scholar 

  • Nakagawa H, Maeda S, Hikiba Y, Ohmae T, Shibata W, Yanai A, Sakamoto K, Ogura K, Noguchi T, Karin M, Ichijo H, Omata M (2008) Deletion of apoptosis signal-regulating kinase 1 attenuates acetaminophen-induced liver injury by inhibition c-Jun N-terminal kinase activation. Gastroenterology 135:1311–1321

    Article  CAS  PubMed  Google Scholar 

  • Ramachandran A, Lebofsky M, Weinman SA, Jaeschke H (2011) The impact of partial manganese superoxide dismutase (SOD2)-deficiency on mitochondrial oxidant stress, DNA fragmentation and liver injury during acetaminophen hepatotoxicity. Toxicol Appl Pharmacol 251:226–233

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Rinaldo P, Matern D, Bennett MJ (2002) Fatty acid oxidation disorders. Annu Rev Physiol 64:477–502

    Article  CAS  PubMed  Google Scholar 

  • Saito C, Lemasters JJ, Jaeschke H (2010a) c-Jun N-terminal kinase modulates oxidant stress and peroxynitrite formation independent of inducible nitric oxide synthase in acetaminophen hepatotoxicity. Toxicol Appl Pharmacol 246:8–17

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Saito C, Zwingmann C, Jaeschke H (2010b) Novel mechanisms of protection against acetaminophen hepatotoxicity in mice by glutathione and N-acetylcysteine. Hepatology 51:246–254

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Sharma M, Gadang V, Jaeschke A (2012) Critical role for mixed-lineage kinase 3 in acetaminophen-induced hepatotoxicity. Mol Pharmacol 82:1001–1007

    Article  CAS  PubMed  Google Scholar 

  • Tirmenstein MA, Nelson SD (1989) Subcellular binding and effects on calcium homeostasis produced by acetaminophen and a nonhepatotoxic regioisomer, 3′-hydroxyacetanilide, in mouse liver. J Biol Chem 264:9814–9819

    CAS  PubMed  Google Scholar 

  • Whitehouse LW, Wong LT, Solomonraj G, Paul CJ, Thomas BH (1981) N-acetylcysteine-induced inhibition of gastric emptying: a mechanism affording protection to mice from the hepatotoxicity of concomitantly administered acetaminophen. Toxicology 19:113–125

    Article  CAS  PubMed  Google Scholar 

  • Win S, Than TA, Han D, Petrovic LM, Kaplowitz N (2011) c-Jun N-terminal kinase (JNK)-dependent acute liver injury from acetaminophen or tumor necrosis factor (TNF) requires mitochondrial Sab protein expression in mice. J Biol Chem 286:35071–35078

    Article  CAS  PubMed  Google Scholar 

  • Wong SG, Card JW, Racz WJ (2000) The role of mitochondrial injury in bromobenzene and furosemide induced hepatotoxicity. Toxicol Lett 116:171–181

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported in part by Grants from McNeil Consumer Health Inc. (to H.J. and S.C.C.), by the University of Kansas Medical Center Liver Center (to H.J.), by the National Institutes of Health Grants R01 DK070195 and R01 AA12916 (to H.J.), and by Grants from the National Center for Research Resources (5P20RR021940-07) and the National Institute of General Medical Sciences (8 P20 GM103549-07) of the National Institutes of Health. Additional support came from an Institutional Development Award (IDeA) from the National Institute of General Medical Sciences of the National Institutes of Health under Grant number P20 GM12345 and, from the “Training Program in Environmental Toxicology” T32 ES007079-26A2 (to M.R.M. and C.D.W.) from the National Institute of Environmental Health Sciences.

Conflict of interest

The authors declare no competing financial interest.

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Authors and Affiliations

  1. Department of Pharmacology, Toxicology, and Therapeutics, University of Kansas Medical Center, 3901 Rainbow Blvd, MS 1018, Kansas City, KS, 66160, USA

    Mitchell R. McGill, Feng Li, C. David Williams, Xiaochao Ma & Hartmut Jaeschke

  2. Department of Internal Medicine, University of Kansas Hospital, Kansas City, KS, USA

    Matthew R. Sharpe

  3. Department of Medical Toxicology, Banner Good Samaritan Medical Center, Phoenix, AZ, USA

    Steven C. Curry

  4. Department of Medicine, Center for Toxicology and Pharmacology Education and Research, University of Arizona College of Medicine, Phoenix, AZ, USA

    Steven C. Curry

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  1. Mitchell R. McGill
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Correspondence to Hartmut Jaeschke.

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McGill, M.R., Li, F., Sharpe, M.R. et al. Circulating acylcarnitines as biomarkers of mitochondrial dysfunction after acetaminophen overdose in mice and humans. Arch Toxicol 88, 391–401 (2014). https://doi.org/10.1007/s00204-013-1118-1

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