title: S-adenosyl-l-methionine protection of acetaminophen mediated oxidative stress and identification of hepatic 4-hydroxynonenal protein adducts by mass spectrometry.
abstract: Acetaminophen (APAP) hepatotoxicity is protected by S-adenosyl-l-methionine (SAMe) treatment 1hour (h) after APAP in C57/Bl6 mice. This study examined protein carbonylation as well as mitochondrial and cytosolic protein adduction by 4-hydroxynonenal (4-HNE) using mass spectrometry (MS) analysis. Additional studies investigated the leakage of mitochondrial proteins and 4-HNE adduction of these proteins. Male C57/Bl6 mice (n=5/group) were divided into the following groups and treated as indicated: Veh (15ml/kg water, ip), SAMe (1.25mmol/kg, ip), APAP (250mg/kg), and SAMe given 1h after APAP (S+A). APAP toxicity was confirmed by an increase (p<0.05) in plasma ALT (U/l) and liver weight/10g body weight relative to the Veh, SAMe and S+A groups 4h following APAP treatment. SAMe administered 1h post-APAP partially corrected APAP hepatotoxicity as ALT and liver weight/10g body weights were lower in the S+A group compared the APAP group. APAP induced leakage of the mitochondrial protein, carbamoyl phosphate synthase-1 (CPS-1) into the cytosol and which was reduced in the S+A group. SAMe further reduced the extent of APAP mediated 4-HNE adduction of CPS-1. MS analysis of hepatic and mitochondrial subcellular fractions identified proteins from APAP treated mice. Site specific 4-HNE adducts were identified on mitochondrial proteins sarcosine dehydrogenase and carbamoyl phosphate synthase-1 (CPS-1). In summary, APAP is associated with 4-HNE adduction of proteins as identified by MS analysis and that CPS-1 leakage was greater in APAP treated mice. SAMe reduced the extent of 4-HNE adduction of proteins as well as leakage of CPS-1.
authors.affiliation: Department of Pharmacology, Physiology and Toxicology, Joan C. Edwards School of Medicine, Huntington, WV, USA.
authors.forename: James Mike
authors.initials: JM
authors.lastname: Brown
authors.affiliation: Southwest Environmental Health Sciences Center, Department of Pharmacology and Toxicology, College of Pharmacy, University of Arizona Health Sciences Center, Tucson, AZ, USA.
authors.forename: Christopher
authors.initials: C
authors.lastname: Kuhlman
authors.affiliation: Department of Pharmacology, Physiology and Toxicology, Joan C. Edwards School of Medicine, Huntington, WV, USA.
authors.forename: Marcus V
authors.initials: MV
authors.lastname: Terneus
authors.affiliation: Southwest Environmental Health Sciences Center, Department of Pharmacology and Toxicology, College of Pharmacy, University of Arizona Health Sciences Center, Tucson, AZ, USA.
authors.forename: Matthew T
authors.initials: MT
authors.lastname: Labenski
authors.affiliation: Department of Pharmacology, Physiology and Toxicology, Joan C. Edwards School of Medicine, Huntington, WV, USA.
authors.forename: Andre Benja
authors.initials: AB
authors.lastname: Lamyaithong
authors.affiliation: Department of Pharmacology, Physiology and Toxicology, Joan C. Edwards School of Medicine, Huntington, WV, USA.
authors.forename: John G
authors.initials: JG
authors.lastname: Ball
authors.affiliation: Southwest Environmental Health Sciences Center, Department of Pharmacology and Toxicology, College of Pharmacy, University of Arizona Health Sciences Center, Tucson, AZ, USA.
authors.forename: Serrine S
authors.initials: SS
authors.lastname: Lau
authors.affiliation: Department of Pharmacology, Physiology and Toxicology, Joan C. Edwards School of Medicine, Huntington, WV, USA. Electronic address: Valentov@marshall.edu.
authors.forename: Monica A
authors.initials: MA
authors.lastname: Valentovic
mesh_terms: D000082:Acetaminophen; D000447:Aldehydes / Q000378:metabolism*; D000818:Animals; D000975:Antioxidants / Q000494:pharmacology*; D002222:Carbamoyl-Phosphate Synthase (Ammonia) / Q000378:metabolism; D056486:Chemical and Drug Induced Liver Injury / Q000209:etiology / Q000378:metabolism / Q000517:prevention & control*; D002853:Chromatography, Liquid; D019610:Cytoprotection; D004195:Disease Models, Animal; D008099:Liver / Q000187:drug effects* / Q000378:metabolism; D008297:Male; D008807:Mice, Inbred BALB C; D008930:Mitochondria, Liver / Q000187:drug effects / Q000378:metabolism; D018384:Oxidative Stress / Q000187:drug effects*; D050050:Protein Carbonylation; D011499:Protein Processing, Post-Translational; D012436:S-Adenosylmethionine / Q000494:pharmacology*; D050892:Sarcosine Dehydrogenase / Q000378:metabolism; D053719:Tandem Mass Spectrometry
chemical_list: D000447:Aldehydes; D000975:Antioxidants; D000082:Acetaminophen; D012436:S-Adenosylmethionine; D050892:Sarcosine Dehydrogenase; C494396:Sardh protein, mouse; D002222:Carbamoyl-Phosphate Synthase (Ammonia); C027576:4-hydroxy-2-nonenal
keywords: 4-Hydroxynonenal; Acetaminophen; Hepatotoxicity; Mass spectrometry; Oxidative stress; Protein adduction