Acetaminophen hepatotoxicity and mechanisms of its protection by N-acetylcysteine: A study of Hep3B cells

Irena Manov, Mark Hirsh, Theodore C. Iancu

Research output: Contribution to journalArticlepeer-review


Acetaminophen (AAP) hepatotoxicity, resulting in centrilobular necrosis, is frequently encountered following suicidal attempts, especially by adolescents, but also after its excessive use in infants. The subcellular and molecular sequences leading to hepatocellular cell death are not yet clear. We therefore investigated AAP hepatotoxicity by using cultured hepatoma-derived cells (Hep3B) exposed to AAP and N-acetylcysteine (NAC), used as a protective agent. Specifically, we studied the role of apoptosis and oxidative damage as putative mechanisms of AAP-associated cytotoxicity. Hep3B cells were exposed to AAP (5-25 mM) and NAC (5 mM) for different time periods. Cell viability was assessed by the Alamar Blue Reduction Test and LDH. Oxidative damage was evaluated by measuring reactive oxygen species (ROS) and glutathione. AAP-induced apoptosis was investigated by flow cytometry and transmission electron microscopy. We found that: 1. In Hep3B cells, AAP causes a time- and concentration-dependent cytotoxic effect, leading to oxidative stress, mitochondrial dysfunction, alterations of membrane permeability and apoptosis; 2. In the course of AAP cytotoxicity, the generation of ROS appears as an early event which precedes decrease of viability, LDH leakage, glutathione depletion and apoptosis; 3. NAC protects Hep3B cells from AAP-induced oxidative injury, but does not prevent apoptosis.

Original languageEnglish
Pages (from-to)489-500
Number of pages12
JournalExperimental and Toxicologic Pathology
Issue number6
StatePublished - 2002
Externally publishedYes

Bibliographical note

Funding Information:
Acknowledgements: The authors wish to thank Mrs. JUDITH REGEV for skillful technical assistance. Dr. Manov was supported in part by the GILADI FUND and by the Mil-man Fund for Pediatric Research (Grant 12-98-99).


  • Acetaminophen
  • Apoptosis
  • Electron microscopy
  • Hep3B cultured cells
  • Hepatotoxicity
  • N-acetylcysteine
  • Oxidative stress
  • Reactive oxygen species

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Toxicology
  • Cell Biology


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