ToxSci Advance Access originally published online on April 28, 2004
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Toxicological Sciences 80, 343-349 (2004)
Toxicological Sciences vol. 80 no. 2 © Society of Toxicology 2004; all rights reserved.
Acetaminophen-Induced Oxidant Stress and Cell Injury in Cultured Mouse Hepatocytes: Protection by N-Acetyl Cysteine
* Liver Research Institute, University of Arizona, College of Medicine, Tucson, Arizona 85724; Department of Pharmacology and Toxicology, University of Arkansas for Medical Sciences, Little Rock, Arkansas 72205; Department of Cell and Developmental Biology, University of North Carolina, Chapel Hill, North Carolina 27599
Received January 13, 2004; accepted April 7, 2004
The increase in cellular and mitochondrial glutathione disulfide (GSSG) levels and the GSSG:GSH ratio after acetaminophen (AAP) overdose suggest the involvement of an oxidant stress in the pathophysiology. However, the initial severe depletion of hepatocellular glutathione makes quantitative assessment of the oxidant stress difficult. Therefore, we tested the hypothesis that oxidant stress precedes the onset of cell injury in a cell culture model using 2',7'-dichlorofluorescein (DCF) fluorescence as a marker for intracellular oxidant stress. Cultured primary murine hepatocytes were exposed to 5 mM AAP. DCF fluorescence, XTT reduction, lactate dehydrogenase (LDH) release, and trypan blue uptake were determined from 0 to 12 h. After glutathione depletion at 3 h, DCF fluorescence increased by 16-fold and was maintained at that level up to 12 h. At 1.5 h after AAP, a significant decrease of the cellular XTT reduction capacity was observed, which continued to decline until 9 h. Cell necrosis (LDH release, trypan blue uptake) was detectable in 20% of cells at 6 h, with a significant further increase at later time points. Pretreatment with 20 mM N-acetylcysteine (NAC) 1 h before AAP enhanced cellular glutathione content, prevented or attenuated the AAP-induced decrease of GSH levels and XTT reduction capacity, respectively, and reduced the loss of cell viability. Additionally, treatment with NAC 2 h after AAP exposure prevented further deterioration of XTT reduction at 3 h and later, and attenuated cell necrosis. Thus, AAP-induced oxidant stress precedes cell necrosis and, in cultured hepatocytes, the oxidant stress is involved in the propagation of cell injury.
Key Words: acetaminophen; hepatotoxicity; oxidant stress; N-acteylcysteine; cultured hepatocytes.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  K. N. Nejak-Bowen, G. Zeng, X. Tan, B. Cieply, and S. P. Monga {beta}-Catenin Regulates Vitamin C Biosynthesis and Cell Survival in Murine Liver J. Biol. Chem., October 9, 2009; 284(41): 28115 - 28127. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. L. Bajt, A. Farhood, J. J. Lemasters, and H. Jaeschke Mitochondrial Bax Translocation Accelerates DNA Fragmentation and Cell Necrosis in a Murine Model of Acetaminophen Hepatotoxicity J. Pharmacol. Exp. Ther., January 1, 2008; 324(1): 8 - 14. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. K. Ramaiah and H. Jaeschke Role of Neutrophils in the Pathogenesis of Acute Inflammatory Liver Injury Toxicol Pathol, October 1, 2007; 35(6): 757 - 766. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. L. Bajt, C. Cover, J. J. Lemasters, and H. Jaeschke Nuclear Translocation of Endonuclease G and Apoptosis-Inducing Factor during Acetaminophen-Induced Liver Cell Injury Toxicol. Sci., November 1, 2006; 94(1): 217 - 225. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J.-H. Zhu and X. G. Lei Double Null of Selenium-Glutathione Peroxidase-1 and Copper, Zinc-Superoxide Dismutase Enhances Resistance of Mouse Primary Hepatocytes to Acetaminophen Toxicity. Experimental Biology and Medicine, May 1, 2006; 231(5): 545 - 552. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Jaeschke and M. L. Bajt Intracellular Signaling Mechanisms of Acetaminophen-Induced Liver Cell Death Toxicol. Sci., January 1, 2006; 89(1): 31 - 41. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Cover, A. Mansouri, T. R. Knight, M. L. Bajt, J. J. Lemasters, D. Pessayre, and H. Jaeschke Peroxynitrite-Induced Mitochondrial and Endonuclease-Mediated Nuclear DNA Damage in Acetaminophen Hepatotoxicity J. Pharmacol. Exp. Ther., November 1, 2005; 315(2): 879 - 887. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Jaeschke Comments on "Glycogen Synthase Kinase-3 Mediates Acetaminophen-Induced Apoptosis in Human Hepatoma Cells" J. Pharmacol. Exp. Ther., September 1, 2005; 314(3): 1401 - 1402. [Full Text] [PDF]
|
|
|
|
|
|
J. Yan and B. F. Hales Activator Protein-1 (AP-1) DNA Binding Activity Is Induced by Hydroxyurea in Organogenesis Stage Mouse Embryos Toxicol. Sci., June 1, 2005; 85(2): 1013 - 1023. [Abstract] [Full Text] [PDF]
|
|