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Archives of Toxicology

, Volume 91, Issue 2, pp 761–773 | Cite as

Mitochondria-targeted antioxidant Mito-Tempo protects against acetaminophen hepatotoxicity

  • Kuo Du
  • Anwar Farhood
  • Hartmut JaeschkeEmail author
Molecular Toxicology

Abstract

Acetaminophen (APAP) hepatotoxicity is characterized by an extensive mitochondrial oxidant stress. However, its importance as a drug target has not been clarified. To investigate this, fasted C57BL/6J mice were treated with 300 mg/kg APAP and the mitochondria-targeted antioxidant Mito-Tempo (MT) was given 1.5 h later. APAP caused severe liver injury in mice, as indicated by the increase in plasma ALT activities and centrilobular necrosis. MT dose-dependently reduced the injury. Importantly, MT did not affect APAP-protein adducts formation, glutathione depletion or c-jun N-terminal kinase activation and its mitochondrial translocation. In contrast, hepatic glutathione disulfide and peroxynitrite formation were dose-dependently reduced by MT, indicating its effective mitochondrial oxidant stress scavenging capacity. Consequently, mitochondrial translocation of Bax and release of mitochondrial intermembrane proteins such as apoptosis-inducing factor were prevented, and nuclear DNA fragmentation was eliminated. To demonstrate the importance of mitochondria-specific antioxidant property of MT, we compared its efficacy with Tempo, which has the same pharmacological mode of action as MT but lacks the mitochondria targeting moiety. In contrast to the dramatic protection by MT, the same molar dose of Tempo did not significantly reduce APAP hepatotoxicity. In contrast, even a 3 h post-treatment with MT reduced 70 % of the injury, and the combination of MT with N-acetylcysteine (NAC) provided superior protection than NAC alone. We conclude that MT protects against APAP overdose in mice by attenuating the mitochondrial oxidant stress and preventing peroxynitrite formation and the subsequent mitochondrial dysfunction. MT is a promising therapeutic agent for APAP overdose patients.

Keywords

Acetaminophen hepatotoxicity Mitochondria Oxidant stress Antioxidant Mito-Tempo 

Abbreviations

AIF

Apoptosis-inducing factor

ALT

Alanine aminotransferase

APAP

Acetaminophen

CYP

Cytochrome P450

GSH

Glutathione

GSSG

Glutathione disulfide

HPLC-ECD

High-pressure liquid chromatography with electrochemical detection

MPT

Mitochondrial permeability transition

MT

Mito-Tempo

NAC

N-acetylcysteine

NAPQI

N-acetyl-p-benzoquinone imine

ROS

Reactive oxygen species

Smac

Second mitochondria-derived activator of caspases

TUNEL

Terminal deoxynucleotidyl transferase dUTP nick end labeling

Notes

Acknowledgments

This investigation was supported in part by the National Institutes of Health grant R01 DK102142 and by Grants from the National Center for Research Resources (5P20RR021940-07) and the National Institute of General Medical Sciences (8 P20 GM103549-07) from the National Institutes of Health. Additional support came an award from the Biomedical Research Training Program (BRTP) from the University of Kansas Medical Center (to K.D.).

Compliance with ethical standards

The manuscript does not contain clinical studies or patient data.

Conflict of interest

The authors declare that there is no conflict of interest.

Supplementary material

204_2016_1692_MOESM1_ESM.docx (17 kb)
Supplementary material 1 (DOCX 17 kb)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  1. 1.Department of Pharmacology, Toxicology and TherapeuticsUniversity of Kansas Medical CenterKansas CityUSA
  2. 2.Department of PathologySt. David’s North Austin Medical CenterAustinUSA

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