Behrends, Volker ORCID: https://orcid.org/0000-0003-4855-5497, Giskeødegård, Guro F., Bravo-Santano, Natalia, Letek, Michal and C. Keun, Hector (2018) Acetaminophen cytotoxicity in HepG2 cells is associated with a decoupling of glycolysis from the TCA cycle, loss of NADPH production, and suppression of anabolism. Archives of Toxicology, 93. ISSN 0340-5761

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Abstract

Acetaminophen (APAP) is one of the most commonly used analgesics worldwide, and overdoses are associated with lactic acidosis, hepatocyte toxicity, and acute liver failure due to oxidative stress and mitochondrial dysfunction. Hepatoma cell lines typically lack the CYP450 activity to generate the reactive metabolite of APAP observed in vivo, but are still subject to APAP cytotoxicity. In this study, we employed metabolic profiling and isotope labelling approaches to investigate the metabolic impact of acute exposure to cytotoxic doses of APAP on the widely used HepG2 cell model. We found that APAP exposure leads to limited cellular death and substantial growth inhibition. Metabolically, we observed an up-regulation of glycolysis and lactate production with a concomitant reduction in carbon from glucose entering the pentose-phosphate pathway and the TCA cycle. This was accompanied by a depletion of cellular NADPH and a reduction in the de novo synthesis of fatty acids and the amino acids serine and glycine. These events were not associated with lower reduced glutathione levels and no glutathione conjugates were seen in cell extracts. Co-treatment with a specific inhibitor of the lactate/H+ transporter MCT1, AZD3965, led to increased apoptosis in APAP-treated cells, suggesting that lactate accumulation could be a cause of cell death in this model. In conclusion, we show that APAP toxicity in HepG2 cells is largely independent of oxidative stress, and is linked instead to a decoupling of glycolysis from the TCA cycle, lactic acidosis, reduced NADPH production, and subsequent suppression of the anabolic pathways required for rapid growth.

Item Type:	Article
Identifier:	10.1007/s00204-018-2371-0
Keywords:	Acetaminophen · Metabolomics · GC–MS · NMR · Isotopomerspectral analysis · HepG2
Subjects:	Natural sciences > Cell and molecular biology
Depositing User:	Volker Behrends
Date Deposited:	29 Apr 2024 09:40
Last Modified:	04 Nov 2024 11:34
URI:	https://repository.uwl.ac.uk/id/eprint/11385

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