Skip to main content
SpringerLink
Log in

The role of calcium in paracetamol (acetaminophen) cytotoxicity in PC12 cells transfected with CYP4502E1

  • Review
  • Published:
InflammoPharmacology Aims and scope Submit manuscript

Abstract

Paracetamol-induced toxicity is mainly due to the accumulation of its CYP450-mediated N-hydroxylation product - N-acetylimidoquinone. We examined cell viability, proliferation rates and intracellular calcium in PC12 cells and in a PC12 cell line transfected with cytochrome P4502E1 exposed to paracetamol. This drug had a concentration-related effect on cell survival and a LD50 which was significantly different between both cell types. A 48% decrease of PC12 cells was found following application of 5 mmol/L paracetamol for 48 h. A total 73% decrease in cell numbers was found in cells metabolizing the drug. Culture protein levels were diminished in a similar manner. Paracetamol increased intracellular calcium (by 662%) only in CYP4502E1-transfected cells. The protective role of EGTA and verapamil modulating calcium homeostasis was more evident in CYP4502E1-transfected cells. These results suggest that biotransformation of paracetamol by CYP2E1 increases its cytotoxicity and that a calcium imbalance may have a key role in the initiation of cell injury.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Insel PA. Analgesic-antipyretic and antiinflammatory agents: drugs employed in the treatment of rheumatoid arthritis and gout. In: Goodman Gilman A, Rall TW, Nies AS, Taylor P, eds. The Pharmacological Basis of Therapeutics. New York: Pergamon Press; 1990:638–81.

    Google Scholar 

  2. Patten CJ, Thomas PE, Guy RL et al. Cytochrome P450 enzymes involved in acetaminophen activation by rat and human liver microsomes and their kinetics. Chem Res Toxicol. 1993;6:511–18.

    Article  PubMed  CAS  Google Scholar 

  3. Holownia A, Mapoles J, Menez JF, Braszko JJ. Acetaminophen metabolism and cytotoxicity in PC12 cells transfected with cytochrome P4502E1. J Mol Med. 1997;75:522–7.

    Article  PubMed  CAS  Google Scholar 

  4. Mapoles J, Berthou F, Alexander A, Simon F, Menez JF. Mammalian PC-12 cell genetically engineered for human cytochrome P450 2E1 expression. Eur J Biochem. 1993;214:735–45.

    Article  PubMed  CAS  Google Scholar 

  5. Day Y, Cederbaum AI. Cytotoxicity of acetaminophen in human cytochrome P4502E1-transfected HepG32 cells. J Pharmacol Exp Ther. 1995;273:1–9.

    Google Scholar 

  6. Faber JL. The role of calcium in lethal cell injury. Chem Res Toxicol. 1990;3:503–8.

    Article  Google Scholar 

  7. Nicotera P, Bellomo G, Orrenius S. The role of Ca2+ in cell killing. Chem Res Toxicol. 1990;3:484–94.

    Article  PubMed  CAS  Google Scholar 

  8. Nicotera P, Bellomo G, Orrenius S. Calcium-mediated mechanisms in chemically induced cell death. Ann Rev Pharmacol Toxicol. 1992;32:449–70.

    Article  CAS  Google Scholar 

  9. Ray SD, Sorge CL, Tavacoli A, Raucy JL, Corcoran GB. Extensive alteration of genomic DNA and rise in nuclear Ca2+ in vivo early after hepatotoxic acetaminophen overdose in mice. Adv Exp Med Biol. 1991;283:699–705.

    PubMed  CAS  Google Scholar 

  10. Shen W, Kamendulis MN, Ray SD, Corcoran GB. Acetaminophen induced cytotoxicity in cultured mouse hepatocytes: correlation of nuclear Ca2+ accumulation and early DNA fragmentation with cell death. Toxicol Appl Pharmacol. 1991;111:245–54.

    Article  Google Scholar 

  11. Arslan P, Di Virgilio F, Beltrame M, Tsien RY, Pozzan T. Cytosolic Ca++ homeostasis in Ehrlich and Yoshida carcinomas. J Biol Chem. 1985;260:2719–27.

    PubMed  CAS  Google Scholar 

  12. Bradford MM. A rapid and sensitive method for the quantitation of the microgram quantities of protein utilizing the principle of protein dye-binding. Anal Biochem. 1976;72:248–54.

    Article  PubMed  CAS  Google Scholar 

  13. Santella L. The cell nucleus: an eldorado to future calcium research. Memb Biol. 1996;153:83–92.

    Article  CAS  Google Scholar 

  14. Masaki N, Kyle ME, Farber JL. Tert-butyl hydroperoxide kills cultured hepatocytes by peroxidizing membrane lipids. Archiv Biochem Biophys. 1989;270:390–9.

    Article  Google Scholar 

  15. Pollard HA, Dessi F, Moreau J, Lasbennes F, BenAri Y, Charriaut-Marlangue C. Regional variability in DNA fragmentation after global cerebral ischemia evidenced by combined histological and gel electrophoresis observations in the rat brain. J Neurochem. 1993;61:1973–6.

    Article  PubMed  Google Scholar 

  16. Ray SD, Kamendulis LM, Gurule MW, Yorkin RD, Corcoran GB. Ca++ antagonists inhibit DNA fragmentation and toxic cell death induced by acetaminophen. FASEB J. 1993;7:453–63.

    PubMed  CAS  Google Scholar 

  17. Barry MA, Reynolds JE, Eastman A. Etoposide-induced apoptosis in human HL-60 cells is associated with intracellular acidification. Cancer Res. 1993;53(Suppl. 10):2349–57.

    PubMed  CAS  Google Scholar 

  18. Ojcius DM, Zychlinski A, Zheng LM, Young JD. Ionophore-induced apoptosis: role of DNA fragmentation and calcium fluxes. Exp Cell Res. 1991;197:43–9.

    Article  PubMed  CAS  Google Scholar 

  19. McConkey DL, Hartzell P, Duddy SK. 2,3,7,8-Tetrachloro-dibenzo-p-dioxin kills immature thymocytes by Ca2+-mediated endonuclease activation. Science. 1988;242:256–9.

    Article  PubMed  CAS  Google Scholar 

  20. Freeman RS, Estus S, Johnson EM. Analysis of cell-related gene expression in postmitotic neurons: selective induction of cyclin D1 during programmed cell death. Neuron. 1994;12:343–55.

    Article  PubMed  CAS  Google Scholar 

  21. Schellmeyer JE. Calpain II involvement in mitosis. Science. 1988;240:911–13.

    Article  Google Scholar 

  22. Squier MKT, Miller ACK, Malkinson AM, Cohen JJ. Calpain activation in apoptosis. J Physiol. 1994;159:229–37.

    CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Medical Academy of Bialystok, Ludwik Zamenhof Children’s Hospital, Clinical Pharmacology Unit, J. Waszyngtona 17, 15-274, Bialystok, Poland

    A. Holownia & J. J. Braszko

  2. Laboratoire de Biochimie-Nutrition, Faculté de Médecine, 22 Avenue Camille Desmoulins, 29285, Brest, France

    J. -F. Menez

Authors
  1. A. Holownia
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. J. -F. Menez
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. J. J. Braszko
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Holownia, A., Menez, J.F. & Braszko, J.J. The role of calcium in paracetamol (acetaminophen) cytotoxicity in PC12 cells transfected with CYP4502E1. Inflammopharmacol 6, 133–142 (1998). https://doi.org/10.1007/s10787-998-0030-4

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10787-998-0030-4

Keywords

Search

Navigation