TY - JOUR
T1 - Hemizygosity of transsulfuration genes confers increased vulnerability against acetaminophen-induced hepatotoxicity in mice
AU - Hagiya, Yoshifumi
AU - Kamata, Shotaro
AU - Mitsuoka, Saya
AU - Okada, Norihiko
AU - Yoshida, Saori
AU - Yamamoto, Junya
AU - Ohkubo, Rika
AU - Abiko, Yumi
AU - Yamada, Hidenori
AU - Akahoshi, Noriyuki
AU - Kasahara, Tadashi
AU - Kumagai, Yoshito
AU - Ishii, Isao
N1 - Funding Information:
This work was supported by the Grants-in-Aid for Scientific Research ( 25460072 and 25220103 ) and the Program for Strategic Research Foundation at Private Universities (2011–2015) from the Ministry of Education, Culture, Sports, Science, and Technology (MEXT) of Japan. We would like to thank Drs. Takehiro Yamamoto and Kyoko Ishiwata (Department of Biochemistry, Keio University School of Medicine) for their advice on 2D DIGE analysis and animal care staff at Keio University School of Pharmaceutical Sciences.
Publisher Copyright:
© 2014 Elsevier Inc.
PY - 2015/1/5
Y1 - 2015/1/5
N2 - 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.
AB - 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.
KW - 2D DIGE
KW - Cystathionine β-synthase
KW - Cystathionine γ-lyase
KW - Glutamate-cysteine ligase
KW - MALDI-TOF/MS
KW - Oncosis
UR - http://www.scopus.com/inward/record.url?scp=84919935008&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84919935008&partnerID=8YFLogxK
U2 - 10.1016/j.taap.2014.11.015
DO - 10.1016/j.taap.2014.11.015
M3 - Article
C2 - 25499718
AN - SCOPUS:84919935008
VL - 282
SP - 195
EP - 206
JO - Toxicology and Applied Pharmacology
JF - Toxicology and Applied Pharmacology
SN - 0041-008X
IS - 2
ER -