Abstract
An overdose of acetaminophen (APAP) causes hepatotoxicity due to its metabolite, N-acetyl-p-benzoquinone imine. NAD(P)H:quinone oxidoreductase 1 (NQO1) is an important enzyme for detoxification, because it catabolizes endogenous/exogenous quinone to hydroquinone. Although various studies have suggested the possible involvement of NQO1 in APAP-induced hepatotoxicity, its precise role in this remains unclear. We investigated the role of NQO1 against APAP-induced hepatotoxicity using a genetically modified rodent model. NQO1 wild-type (WT) and knockout (KO) mice were treated with different doses of APAP, and we evaluated the mortality and toxicity markers for cell death caused by APAP. NQO1 KO mice showed high sensitivity to APAP-mediated hepatotoxicity (as indicated by a large necrotic region) as well as increased levels of nitrotyrosine adducts and reactive oxygen species. APAP-induced cell death in the livers and primary hepatocytes of NQO1 KO mice, which was accompanied by an extensive reduction in adenosine triphosphate (ATP) levels. In accordance with this ATP depletion, cytosolic increases in mitochondrial proteins such as apoptosis-inducing factor, second mitochondria-derived activator of caspases/DIABLO, endonuclease G, and cytochrome c, which indicate severe mitochondrial dysfunction, were observed in NQO1 KO mice but not in WT mice after APAP exposure. Severe mitochondrial depolarization was also greater in hepatocytes isolated from NQO1 KO mice. Collectively, our data suggest that NQO1 plays a critical role in protection against energy depletion caused by APAP, and NQO1 may be useful in the development of therapeutic approaches to effectively diminish the hepatotoxicity caused by an APAP overdose.
Abbreviations
- NQO1:
-
NAD(P)H:quinone oxidoreductase
- APAP:
-
Acetaminophen
- NAPQI:
-
N-acetyl-p-benzoquinone imine
- ROS:
-
Reactive oxygen species
- GSH:
-
Glutathione
- MPT:
-
Mitochondrial permeability transition
- ALT:
-
Alanine aminotransferase
- AST:
-
Aspartate aminotransferase
- AIF:
-
Apoptosis-inducing factor
- Endo G:
-
Endonuclease G
- RNS:
-
Reactive nitrogen species
- JNK:
-
c-Jun N-terminal kinase
- ETC:
-
Electron transport chain
- PCNA:
-
Proliferating cell nuclear antigen
- ESI:
-
Electrospray ionization
References
Aleksunes LM, Goedken M, Manautou JE (2006a) Up-regulation of NAD(P)H quinone oxidoreductase 1 during human liver injury. World J Gastroenterol 12:1937–1940
Aleksunes LM, Slitt AL, Maher JM et al (2006b) Nuclear factor-E2-related factor 2 expression in liver is critical for induction of NAD(P)H:quinone oxidoreductase 1 during cholestasis. Cell Stress Chaperones 11:356–363
Ben-Shachar D, Zuk R, Glinka Y (1995) Dopamine neurotoxicity: inhibition of mitochondrial respiration. J Neurochem 64:718–723
Burcham PC, Harman AW (1991) Acetaminophen toxicity results in site-specific mitochondrial damage in isolated mouse hepatocytes. J Biol Chem 266:5049–5054
Cohen SD, Khairallah EA (1997) Selective protein arylation and acetaminophen-induced hepatotoxicity. Drug Metab Rev 29:59–77
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 U S A 81:1327–1331
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. doi:10.1053/j.gastro.2006.03.045
Harman AW, Kyle ME, Serroni A, Farber JL (1991) The killing of cultured hepatocytes by N-acetyl-p-benzoquinone imine (NAPQI) as a model of the cytotoxicity of acetaminophen. Biochem Pharmacol 41:1111–1117
Hwang JH, Kim DW, Jo EJ et al (2009) Pharmacological stimulation of NADH oxidation ameliorates obesity and related phenotypes in mice. Diabetes 58:965–974. doi:10.2337/db08-1183
Jaeschke H, Bajt ML (2006) Intracellular signaling mechanisms of acetaminophen-induced liver cell death. Toxicol Sci 89:31–41. doi:10.1093/toxsci/kfi336
Kim YH, Hwang JH, Noh JR et al (2011) Activation of NAD(P)H:quinone oxidoreductase ameliorates spontaneous hypertension in an animal model via modulation of eNOS activity. Cardiovasc Res 91:519–527
Kim YH, Hwang JH, Noh JR et al (2012) Prevention of salt-induced renal injury by activation of NAD(P)H:quinone oxidoreductase 1, associated with NADPH oxidase. Free Radic Biol Med 52:880–888. doi:10.1016/j.freeradbiomed.2011.12.007
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. doi:10.1002/hep.20437
Larson AM, Polson J, Fontana RJ et al (2005) Acetaminophen-induced acute liver failure: results of a United States multicenter, prospective study. Hepatology 42:1364–1372. doi:10.1002/hep.20948
Lee SM, Cho TS, Kim DJ, Cha YN (1999) Protective effect of ethanol against acetaminophen-induced hepatotoxicity in mice: role of NADH:quinone reductase. Biochem Pharmacol 58:1547–1555
Moffit JS, Aleksunes LM, Kardas MJ, Slitt AL, Klaassen CD, Manautou JE (2007) Role of NAD(P)H:quinone oxidoreductase 1 in clofibrate-mediated hepatoprotection from acetaminophen. Toxicology 230:197–206. doi:10.1016/j.tox.2006.11.052
Nelson SD (1990) Molecular mechanisms of the hepatotoxicity caused by acetaminophen. Semin Liver Dis 10:267–278. doi:10.1055/s-2008-1040482
Pelicano H, Feng L, Zhou Y et al (2003) Inhibition of mitochondrial respiration: a novel strategy to enhance drug-induced apoptosis in human leukemia cells by a reactive oxygen species-mediated mechanism. J Biol Chem 278:37832–37839. doi:10.1074/jbc.M301546200
Prescott LF (1983) Paracetamol overdosage. Pharmacological considerations and clinical management. Drugs 25:290–314
Ross D (2004) Quinone reductases multitasking in the metabolic world. Drug Metab Rev 36:639–654. doi:10.1081/DMR-200033465
Ross D, Kepa JK, Winski SL, Beall HD, Anwar A, Siegel D (2000) NAD(P)H:quinone oxidoreductase 1 (NQO1): chemoprotection, bioactivation, gene regulation and genetic polymorphisms. Chem Biol Interact 129:77–97
Rundgren M, Porubek DJ, Harvison PJ, Cotgreave IA, Moldeus P, Nelson SD (1988) Comparative cytotoxic effects of N-acetyl-p-benzoquinone imine and two dimethylated analogues. Mol Pharmacol 34:566–572
Vercesi AE, Kowaltowski AJ, Grijalba MT, Meinicke AR, Castilho RF (1997) The role of reactive oxygen species in mitochondrial permeability transition. Biosci Rep 17:43–52
Yeo SH, Noh JR, Kom YH et al (2013) Increased vulnerability to beta-cell destruction and diabetes in mice lacking NAD(P)H:quinone oxidoreductase 1. Toxicol Lett 219:35–41. doi:10.1016/j.toxlet.2013.02.013
Acknowledgments
This work was supported by the KRIBB Research Initiative Program of Korea and a grant from the Korea Healthcare Technology R&D Project, Ministry for Health, Welfare, and Family Affairs, Republic of Korea (H10C0573), and a grant from the National Research Foundation of Korea (NRF) funded by the Korean government (NRF-2013R1A2A1A01011071). The authors thank Dong-Hee Choi for technical assistance.
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The authors declare that there are no conflict of interest.
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Jung Hwan Hwang and Yong-Hoon Kim have contributed equally to this work.
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Hwang, J.H., Kim, YH., Noh, JR. et al. The protective role of NAD(P)H:quinone oxidoreductase 1 on acetaminophen-induced liver injury is associated with prevention of adenosine triphosphate depletion and improvement of mitochondrial dysfunction. Arch Toxicol 89, 2159–2166 (2015). https://doi.org/10.1007/s00204-014-1340-5
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DOI: https://doi.org/10.1007/s00204-014-1340-5