Document Detail


Acetaminophen-induced cytotoxicity in cultured mouse hepatocytes: effects of Ca(2+)-endonuclease, DNA repair, and glutathione depletion inhibitors on DNA fragmentation and cell death.
MedLine Citation:
PMID:  1310169     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Hepatotoxic alkylation of mouse liver cells by acetaminophen is characterized by an early loss of ion regulation, accumulation of Ca2+ in the nucleus, and fragmentation of DNA in vitro and in vivo. Acetaminophen-induced DNA cleavage is accompanied by the formation of a "ladder" of DNA fragments characteristic of Ca(2+)-mediated endonuclease activation. These events unfold well in advance of cytotoxicity and the development of necrosis. The present study utilized cultured mouse hepatocytes and mechanistic probes to test whether DNA fragmentation and cell death might be related in a "cause-and-effect" manner. Cells were isolated by collagenase perfusion, cultured in Williams' E medium for 22-26 hr, and exposed to acetaminophen. Aurintricarboxylic acid, a general Ca(2+)-endonuclease inhibitor, and EGTA, a chelator of Ca2+ required for endonuclease activation, significantly decreased DNA fragmentation at 6 and 12 hr and virtually abolished cytotoxicity. N-Acetylcysteine also eliminated DNA fragmentation and cytotoxicity. 3-Aminobenzamide, an inhibitor of poly(ADP-ribose) polymerase-stimulated DNA repair, failed to alter the amount of DNA fragmentation at 6 hr but substantially increased acetaminophen cytotoxicity in hepatocytes at 12 hr. With the exception of when DNA repair was inhibited by 3-aminobenzamide, Ca2+ accumulation in the nucleus, DNA fragmentation, and hepatocyte death varied consistently and predictably with one another. Collectively, these findings suggest that unrepaired damage to DNA contributes to acetaminophen-induced cell death in vivo and may play a role in necrosis in vivo.
Authors:
W Shen; L M Kamendulis; S D Ray; G B Corcoran
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Toxicology and applied pharmacology     Volume:  112     ISSN:  0041-008X     ISO Abbreviation:  Toxicol. Appl. Pharmacol.     Publication Date:  1992 Jan 
Date Detail:
Created Date:  1992-02-27     Completed Date:  1992-02-27     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  0416575     Medline TA:  Toxicol Appl Pharmacol     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  32-40     Citation Subset:  IM    
Affiliation:
Toxicology Program, College of Pharmacy, University of New Mexico, Albuquerque 87131-1066.
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MeSH Terms
Descriptor/Qualifier:
Acetaminophen / adverse effects*
Acetylcysteine / pharmacology
Adenosine Diphosphate / metabolism
Animals
Aurintricarboxylic Acid / pharmacology
Benzamides / pharmacology
Calcium / metabolism*,  pharmacology
Cell Death / drug effects,  physiology
Cell Nucleus / drug effects,  metabolism
Cells, Cultured
DNA / drug effects*,  metabolism
DNA Damage*
DNA Repair*
Egtazic Acid / pharmacology
Endonucleases / drug effects,  metabolism*,  physiology
Enzyme Activation / drug effects
Glutathione / metabolism*
Liver / cytology*,  drug effects
Male
Mice
Grant Support
ID/Acronym/Agency:
GM 41564/GM/NIGMS NIH HHS
Chemical
Reg. No./Substance:
0/Benzamides; 103-90-2/Acetaminophen; 3544-24-9/3-aminobenzamide; 4431-00-9/Aurintricarboxylic Acid; 58-64-0/Adenosine Diphosphate; 616-91-1/Acetylcysteine; 67-42-5/Egtazic Acid; 70-18-8/Glutathione; 7440-70-2/Calcium; 9007-49-2/DNA; EC 3.1.-/Endonucleases

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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