![[Feedback]](/icons/banner/feedbackACT.gif){kind=icon}
![[For Subscribers]](/icons/banner/subscriberACT.gif){kind=icon}
![[Archive]](/icons/banner/archiveACT.gif){kind=icon}
![[Search]](/icons/banner/searchACT.gif){kind=icon}
|
|
|
|
|
||
| |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Received for publication August 10, 2004.
Revised September 30, 2004.
Accepted for publication October 1, 2004.
Freshly isolated mouse hepatocytes were used to determine the role of mitochondrial permeability transition (MPT) in APAP toxicity. Incubation of APAP (1mM) with hepatocytes resulted in cell death as indicated by increased ALT and propidium iodide fluorescence. To separate metabolic events from later events in toxicity, hepatocytes were preincubated with APAP for 2 hr followed by centrifugations of the cells and resuspension of the pellet to remove the drug, and reincubating the cells in media alone. At 2 hr toxicity was not significantly different between control and APAP incubated cells; however, preincubation with APAP followed by reincubation with media alone resulted in a marked increase in toxicity at 3 to 5 hr that was not different from incubation with APAP for the entire time. Inclusion of cyclosporine A, trifluoperazine, dithiothreitol (DTT), or N-acetylcysteine (NAC) in the reincubation phase prevented hepatocyte toxicity. Dichlorofluorescein fluorescence increased during the reincubation phase indicating increased oxidative stress. TMRM fluorescence decreased during the reincubation phase indicating a loss of mitochondrial membrane potential. Inclusion of cyclosporine A, DTT, or NAC decreased oxidative stress and loss of mitochondrial membrane potential. Confocal microscopy studies with the dye calcein AM indicated that MPT had also occurred. These data are consistent with a hypothesis where APAP-induced cell death occurs by two phases, a metabolic phase and an oxidative phase. The metabolic phase occurs with GSH depletion and APAP- protein binding. The oxidative phase occurs with increased oxidative stress, loss of mitochondrial membrane potential, MPT, and toxicity.
Key words:
acetaminophen, hepatocytes, hepatotoxicity, mitochondrial permeability transition, necrosis, oxidative stress
This article has been cited by other articles:
|
|
 |
|
  V. Koshkin, F. F. Dai, C. A. Robson-Doucette, C. B. Chan, and M. B. Wheeler Limited Mitochondrial Permeabilization Is an Early Manifestation of Palmitate-induced Lipotoxicity in Pancreatic {beta}-Cells J. Biol. Chem., March 21, 2008; 283(12): 7936 - 7948. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Chen, K. W. Krausz, J. R. Idle, and F. J. Gonzalez Identification of Novel Toxicity-associated Metabolites by Metabolomics and Mass Isotopomer Analysis of Acetaminophen Metabolism in Wild-type and Cyp2e1-null Mice J. Biol. Chem., February 22, 2008; 283(8): 4543 - 4559. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. A. McConnachie, I. Mohar, F. N. Hudson, C. B. Ware, W. C. Ladiges, C. Fernandez, S. Chatterton-Kirchmeier, C. C. White, R. H. Pierce, and T. J. Kavanagh Glutamate Cysteine Ligase Modifier Subunit Deficiency and Gender as Determinants of Acetaminophen-Induced Hepatotoxicity in Mice Toxicol. Sci., October 1, 2007; 99(2): 628 - 636. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. Pacher, J. S. Beckman, and L. Liaudet Nitric Oxide and Peroxynitrite in Health and Disease Physiol Rev, January 1, 2007; 87(1): 315 - 424. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J.-H. Zhu, X. Zhang, J. P. McClung, and X. G. Lei Impact of Cu,Zn-Superoxide Dismutase and Se-Dependent Glutathione Peroxidase-1 Knockouts on Acetaminophen-Induced Cell Death and Related Signaling in Murine Liver Experimental Biology and Medicine, December 1, 2006; 231(11): 1726 - 1732. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. Alirol, D. James, D. Huber, A. Marchetto, L. Vergani, J.-C. Martinou, and L. Scorrano The Mitochondrial Fission Protein hFis1 Requires the Endoplasmic Reticulum Gateway to Induce Apoptosis Mol. Biol. Cell, November 1, 2006; 17(11): 4593 - 4605. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. Botta, S. Shi, C. C. White, M. J. Dabrowski, C. L. Keener, S. L. Srinouanprachanh, F. M. Farin, C. B. Ware, W. C. Ladiges, R. H. Pierce, et al. Acetaminophen-induced Liver Injury Is Attenuated in Male Glutamate-cysteine Ligase Transgenic Mice J. Biol. Chem., September 29, 2006; 281(39): 28865 - 28875. [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]
|
|
|
|
|
|
V. Tong, X. W. Teng, T. K. H. Chang, and F. S. Abbott Valproic Acid II: Effects on Oxidative Stress, Mitochondrial Membrane Potential, and Cytotoxicity in Glutathione-Depleted Rat Hepatocytes Toxicol. Sci., August 1, 2005; 86(2): 436 - 443. [Abstract] [Full Text] [PDF]
|
|
 |
 |
 |
|
 |
 |
 |
