skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Hemizygosity of transsulfuration genes confers increased vulnerability against acetaminophen-induced hepatotoxicity in mice

Abstract

The key mechanism for acetaminophen hepatotoxicity is cytochrome P450 (CYP)-dependent formation of N-acetyl-p-benzoquinone imine, a potent electrophile that forms protein adducts. Previous studies revealed the fundamental role of glutathione, which binds to and detoxifies N-acetyl-p-benzoquinone imine. Glutathione is synthesized from cysteine in the liver, and N-acetylcysteine is used as a sole antidote for acetaminophen poisoning. Here, we evaluated the potential roles of transsulfuration enzymes essential for cysteine biosynthesis, cystathionine β-synthase (CBS) and cystathionine γ-lyase (CTH), in acetaminophen hepatotoxicity using hemizygous (Cbs{sup +/−} or Cth{sup +/−}) and homozygous (Cth{sup −/−}) knockout mice. At 4 h after intraperitoneal acetaminophen injection, serum alanine aminotransferase levels were highly elevated in Cth{sup −/−} mice at 150 mg/kg dose, and also in Cbs{sup +/−} or Cth{sup +/−} mice at 250 mg/kg dose, which was associated with characteristic centrilobular hepatocyte oncosis. Hepatic glutathione was depleted while serum malondialdehyde accumulated in acetaminophen-injected Cth{sup −/−} mice but not wild-type mice, although glutamate–cysteine ligase (composed of catalytic [GCLC] and modifier [GCLM] subunits) became more activated in the livers of Cth{sup −/−} mice with lower K{sub m} values for Cys and Glu. Proteome analysis using fluorescent two-dimensional difference gel electrophoresis revealed 47 differentially expressed proteins after injection of 150 mg acetaminophen/kgmore » into Cth{sup −/−} mice; the profiles were similar to 1000 mg acetaminophen/kg-treated wild-type mice. The prevalence of Cbs or Cth hemizygosity is estimated to be 1:200–300 population; therefore, the deletion or polymorphism of either transsulfuration gene may underlie idiosyncratic acetaminophen vulnerability along with the differences in Cyp, Gclc, and Gclm gene activities. - Highlights: • Cbs{sup +/−}, Cth{sup +/−}, and especially Cth{sup −/−} mice were susceptible to APAP hepatic injury. • Hepatic glutathione became rapidly depleted upon APAP injection in Cth{sup −/−} mice. • Hepatic glutamate–cysteine ligase was activated by APAP injection and CTH deletion. • 2D DIGE identified 47 differentially expressed hepatic proteins by APAP injection. • Both transsulfuration enzymes are essential for protection against APAP injury.« less

Authors:
; ; ; ; ; ;  [1];  [2];  [3];  [4];  [1];  [2];  [1]
  1. Department of Biochemistry, Keio University School of Pharmaceutical Sciences, Tokyo 105-8512 (Japan)
  2. Environmental Biology Laboratory, School of Medicine, University of Tsukuba, Ibaraki 305-8575 (Japan)
  3. Jobu Hospital for Respiratory Diseases, Maebashi 371-0048 (Japan)
  4. Department of Immunology, Akita University Graduate School of Medicine, Akita 010-8543 (Japan)
Publication Date:
OSTI Identifier:
22465682
Resource Type:
Journal Article
Resource Relation:
Journal Name: Toxicology and Applied Pharmacology; Journal Volume: 282; Journal Issue: 2; Other Information: Copyright (c) 2014 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved.; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; ADDUCTS; ALANINES; BIOSYNTHESIS; CYSTEINE; ELECTROPHORESIS; FLUORESCENCE; GELS; GENES; GLUTATHIONE; GUANINE; INJECTION; INJURIES; LIGASES; LIVER; MICE; POISONING; SUPEROXIDE DISMUTASE; TWO-DIMENSIONAL CALCULATIONS; VULNERABILITY

Citation Formats

Hagiya, Yoshifumi, Kamata, Shotaro, Mitsuoka, Saya, Okada, Norihiko, Yoshida, Saori, Yamamoto, Junya, Ohkubo, Rika, Abiko, Yumi, Yamada, Hidenori, Akahoshi, Noriyuki, Kasahara, Tadashi, Kumagai, Yoshito, and Ishii, Isao, E-mail: isao-ishii@umin.ac.jp. Hemizygosity of transsulfuration genes confers increased vulnerability against acetaminophen-induced hepatotoxicity in mice. United States: N. p., 2015. Web. doi:10.1016/J.TAAP.2014.11.015.
Hagiya, Yoshifumi, Kamata, Shotaro, Mitsuoka, Saya, Okada, Norihiko, Yoshida, Saori, Yamamoto, Junya, Ohkubo, Rika, Abiko, Yumi, Yamada, Hidenori, Akahoshi, Noriyuki, Kasahara, Tadashi, Kumagai, Yoshito, & Ishii, Isao, E-mail: isao-ishii@umin.ac.jp. Hemizygosity of transsulfuration genes confers increased vulnerability against acetaminophen-induced hepatotoxicity in mice. United States. doi:10.1016/J.TAAP.2014.11.015.
Hagiya, Yoshifumi, Kamata, Shotaro, Mitsuoka, Saya, Okada, Norihiko, Yoshida, Saori, Yamamoto, Junya, Ohkubo, Rika, Abiko, Yumi, Yamada, Hidenori, Akahoshi, Noriyuki, Kasahara, Tadashi, Kumagai, Yoshito, and Ishii, Isao, E-mail: isao-ishii@umin.ac.jp. Thu . "Hemizygosity of transsulfuration genes confers increased vulnerability against acetaminophen-induced hepatotoxicity in mice". United States. doi:10.1016/J.TAAP.2014.11.015.
@article{osti_22465682,
title = {Hemizygosity of transsulfuration genes confers increased vulnerability against acetaminophen-induced hepatotoxicity in mice},
author = {Hagiya, Yoshifumi and Kamata, Shotaro and Mitsuoka, Saya and Okada, Norihiko and Yoshida, Saori and Yamamoto, Junya and Ohkubo, Rika and Abiko, Yumi and Yamada, Hidenori and Akahoshi, Noriyuki and Kasahara, Tadashi and Kumagai, Yoshito and Ishii, Isao, E-mail: isao-ishii@umin.ac.jp},
abstractNote = {The key mechanism for acetaminophen hepatotoxicity is cytochrome P450 (CYP)-dependent formation of N-acetyl-p-benzoquinone imine, a potent electrophile that forms protein adducts. Previous studies revealed the fundamental role of glutathione, which binds to and detoxifies N-acetyl-p-benzoquinone imine. Glutathione is synthesized from cysteine in the liver, and N-acetylcysteine is used as a sole antidote for acetaminophen poisoning. Here, we evaluated the potential roles of transsulfuration enzymes essential for cysteine biosynthesis, cystathionine β-synthase (CBS) and cystathionine γ-lyase (CTH), in acetaminophen hepatotoxicity using hemizygous (Cbs{sup +/−} or Cth{sup +/−}) and homozygous (Cth{sup −/−}) knockout mice. At 4 h after intraperitoneal acetaminophen injection, serum alanine aminotransferase levels were highly elevated in Cth{sup −/−} mice at 150 mg/kg dose, and also in Cbs{sup +/−} or Cth{sup +/−} mice at 250 mg/kg dose, which was associated with characteristic centrilobular hepatocyte oncosis. Hepatic glutathione was depleted while serum malondialdehyde accumulated in acetaminophen-injected Cth{sup −/−} mice but not wild-type mice, although glutamate–cysteine ligase (composed of catalytic [GCLC] and modifier [GCLM] subunits) became more activated in the livers of Cth{sup −/−} mice with lower K{sub m} values for Cys and Glu. Proteome analysis using fluorescent two-dimensional difference gel electrophoresis revealed 47 differentially expressed proteins after injection of 150 mg acetaminophen/kg into Cth{sup −/−} mice; the profiles were similar to 1000 mg acetaminophen/kg-treated wild-type mice. The prevalence of Cbs or Cth hemizygosity is estimated to be 1:200–300 population; therefore, the deletion or polymorphism of either transsulfuration gene may underlie idiosyncratic acetaminophen vulnerability along with the differences in Cyp, Gclc, and Gclm gene activities. - Highlights: • Cbs{sup +/−}, Cth{sup +/−}, and especially Cth{sup −/−} mice were susceptible to APAP hepatic injury. • Hepatic glutathione became rapidly depleted upon APAP injection in Cth{sup −/−} mice. • Hepatic glutamate–cysteine ligase was activated by APAP injection and CTH deletion. • 2D DIGE identified 47 differentially expressed hepatic proteins by APAP injection. • Both transsulfuration enzymes are essential for protection against APAP injury.},
doi = {10.1016/J.TAAP.2014.11.015},
journal = {Toxicology and Applied Pharmacology},
number = 2,
volume = 282,
place = {United States},
year = {Thu Jan 15 00:00:00 EST 2015},
month = {Thu Jan 15 00:00:00 EST 2015}
}