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

Yoshifumi Hagiya, Shotaro Kamata, Saya Mitsuoka, Norihiko Okada, Saori Yoshida, Junya Yamamoto, Rika Ohkubo, Yumi Abiko, Hidenori Yamada, Noriyuki Akahoshi, Tadashi Kasahara, Yoshito Kumagai, Isao Ishii

Research output: Contribution to journalArticle

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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+/- or Cth+/-) and homozygous (Cth-/-) knockout mice. At 4h after intraperitoneal acetaminophen injection, serum alanine aminotransferase levels were highly elevated in Cth-/- mice at 150mg/kg dose, and also in Cbs+/- or Cth+/- mice at 250mg/kg dose, which was associated with characteristic centrilobular hepatocyte oncosis. Hepatic glutathione was depleted while serum malondialdehyde accumulated in acetaminophen-injected Cth-/- 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-/- mice with lower Km values for Cys and Glu. Proteome analysis using fluorescent two-dimensional difference gel electrophoresis revealed 47 differentially expressed proteins after injection of 150mgacetaminophen/kg into Cth-/- mice; the profiles were similar to 1000mgacetaminophen/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.

Original languageEnglish
Pages (from-to)195-206
Number of pages12
JournalToxicology and Applied Pharmacology
Volume282
Issue number2
DOIs
Publication statusPublished - 2015 Jan 5

Fingerprint

Acetaminophen
Genes
Cystathionine
Glutathione
Imines
Liver
Cysteine
Glutamate-Cysteine Ligase
Two-Dimensional Difference Gel Electrophoresis
Antidotes
Lyases
Biosynthesis
Acetylcysteine
Proteome
Electrophoresis
Malondialdehyde
Polymorphism
Alanine Transaminase
Cytochrome P-450 Enzyme System
Intraperitoneal Injections

Keywords

  • 2D DIGE
  • Cystathionine β-synthase
  • Cystathionine γ-lyase
  • Glutamate-cysteine ligase
  • MALDI-TOF/MS
  • Oncosis

ASJC Scopus subject areas

  • Pharmacology
  • Toxicology
  • Medicine(all)

Cite this

Hemizygosity of transsulfuration genes confers increased vulnerability against acetaminophen-induced hepatotoxicity in mice. / 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.

In: Toxicology and Applied Pharmacology, Vol. 282, No. 2, 05.01.2015, p. 195-206.

Research output: Contribution to journalArticle

Hagiya, Y, Kamata, S, Mitsuoka, S, Okada, N, Yoshida, S, Yamamoto, J, Ohkubo, R, Abiko, Y, Yamada, H, Akahoshi, N, Kasahara, T, Kumagai, Y & Ishii, I 2015, 'Hemizygosity of transsulfuration genes confers increased vulnerability against acetaminophen-induced hepatotoxicity in mice', Toxicology and Applied Pharmacology, vol. 282, no. 2, pp. 195-206. https://doi.org/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. / Hemizygosity of transsulfuration genes confers increased vulnerability against acetaminophen-induced hepatotoxicity in mice. In: Toxicology and Applied Pharmacology. 2015 ; Vol. 282, No. 2. pp. 195-206.
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