title: Lack of Direct Cytotoxicity of Extracellular ATP against Hepatocytes: Role in the Mechanism of Acetaminophen Hepatotoxicity.
abstract: BACKGROUND
  Acetaminophen (APAP) hepatotoxicity is a major cause of acute liver failure in many countries. Mechanistic studies in mice and humans have implicated formation of a reactive metabolite, mitochondrial dysfunction and oxidant stress as critical events in the pathophysiology of APAP-induced liver cell death. It was recently suggested that ATP released from necrotic cells can directly cause cell death in mouse hepatocytes and in a hepatoma cell line (HepG2).
  
  
  OBJECTIVE
  To assess if ATP can directly cause cell toxicity in hepatocytes and evaluate their relevance in the human system.
  
  
  METHODS
  Primary mouse hepatocytes, human HepG2 cells, the metabolically competent human HepaRG cell line and freshly isolated primary human hepatocytes were exposed to 10-100 μM ATP or ATγP in the presence or absence of 5-10 mM APAP for 9-24 h.
  
  
  RESULTS
  ATP or ATγP was unable to directly cause cell toxicity in all 4 types of hepatocytes. In addition, ATP did not enhance APAP-induced cell death observed in primary mouse or human hepatocytes, or in HepaRG cells as measured by LDH release and by propidium iodide staining in primary mouse hepatocytes. Furthermore, addition of ATP did not cause mitochondrial dysfunction or enhance APAP-induced mitochondrial dysfunction in primary murine hepatocytes, although ATP did cause cell death in murine RAW macrophages.
  
  
  CONCLUSIONS
  It is unlikely that ATP released from necrotic cells can significantly affect cell death in human or mouse liver during APAP hepatotoxicity.
  Understanding the mechanisms of APAP-induced cell injury is critical for identifying novel therapeutic targets to prevent liver injury and acute liver failure in APAP overdose patients.
authors.affiliation: Department of Pharmacology, Toxicology and Therapeutics, University of Kansas Medical Center, Kansas City, KS 66160, USA.
authors.forename: Yuchao
authors.initials: Y
authors.lastname: Xie
authors.affiliation: Department of Pharmacology, Toxicology and Therapeutics, University of Kansas Medical Center, Kansas City, KS 66160, USA.
authors.forename: Benjamin L
authors.initials: BL
authors.lastname: Woolbright
authors.affiliation: Department of Experimental Surgery, Academic Medical Center, University of Amsterdam, University of Amsterdam, 1105 AZ Amsterdam, The Netherlands.
authors.forename: Milan
authors.initials: M
authors.lastname: Kos
authors.affiliation: Department of Pharmacology, Toxicology and Therapeutics, University of Kansas Medical Center, Kansas City, KS 66160, USA.
authors.forename: Mitchell R
authors.initials: MR
authors.lastname: McGill
authors.affiliation: Department of Pharmacology, Toxicology and Therapeutics, University of Kansas Medical Center, Kansas City, KS 66160, USA.
authors.forename: Kenneth
authors.initials: K
authors.lastname: Dorko
authors.affiliation: Department of Surgery, University of Kansas Medical Center, Kansas City, KS 66160, USA.
authors.forename: Sean C
authors.initials: SC
authors.lastname: Kumer
authors.affiliation: Department of Surgery, University of Kansas Medical Center, Kansas City, KS 66160, USA.
authors.forename: Timothy M
authors.initials: TM
authors.lastname: Schmitt
authors.affiliation: Department of Pharmacology, Toxicology and Therapeutics, University of Kansas Medical Center, Kansas City, KS 66160, USA.
authors.forename: Hartmut
authors.initials: H
authors.lastname: Jaeschke
keywords: Acetaminophen hepatotoxicity; damage-associated molecular patterns; human hepatocytes; necrosis; sterile inflammation