Skip to main content

Menadione-induced cytotoxicity in rat platelets: Absence of the detoxifying enzyme, quinone reductase

Abstract

The elevation of intracellular Ca2+ in various tissue through oxidative stress induced by menadione has been well documented. Increase of Ca2+ level in platelets results in aggregation of platelets. To test the hypothesis that menadione-induced Ca2+ elevations can play a role in platelet aggregation, we have studied the effect of menadione on aggregation of platelets isolated from female rats. Treatment with menadione to platelet rich plasma (PRP), which proved to be an adequate system, appeared to induce dose-dependent turbidity changes of platelets up to 60%, as determined by aggregometry. However, exposure of PRP to menadione leads to a loss of cell viability, as measured by lactate dehydrogenase (LDH) leakage, suggesting that menadione might induce cell lysis rather than aggregation of platelets. Turbidity changes induced by menadione were unaffected by addition of dicoumarol, which is a quinone reductase (QR) inhibitor. Consistent with these findings, no activity of QR was detected in any subcellular fractions of platelets. These data, which indicate an absence of the QR detoxifying pathway, suggest that platelets may be more susceptible to menadione-induced cytotoxicity than certain other cell, such as hepatocytes.

This is a preview of subscription content, access via your institution.

References Cited

  1. Benson, A. M., Hunkeler, M. J. and Talalay, A. P., Increase of NAD(P)H guinone reductase by dietary antioxidants: possible role in protection against carcinogenesis and toxicity.Proc. Natl. Acad. Sci. USA 77, 5216–5220 (1980).

    PubMed  Article  CAS  Google Scholar 

  2. Brass, L. F., Ca+2 homeostasis in unstimulated platelet.J. Biol. Chem., 259, 12563–12570 (1984).

    PubMed  CAS  Google Scholar 

  3. Dicker, E. and Cederbaum, A. I., Requirement for iron for the production of hydroxyl radicals by rat liver quinone reductase.J. Pharmacol. Exp. Ther. 266, 1282–1290 (1993).

    PubMed  CAS  Google Scholar 

  4. Di Monte, D., Ross, D., Bellomo, G., Eklow, L. and Orrenius, S., Alterations in intracellular thiol homeostasis during the metabolism of menadione isolated rat hepatocyte.Arch. Biochem. Biochem. Biophys. 235, 334–342 (1984a).

    Article  Google Scholar 

  5. Di Monte, D., Bellomo, G., Thor, H., Nicotera, P. and Orenius, S., Menadione induced cytotoxicity is associated with thiol oxidation and alteration in intracellular Ca+2 homeostasis.Arch. Biochem. Biophys. 235, 343–350 (1984b).

    PubMed  Article  Google Scholar 

  6. Frojmovic, M. M. and Milton, J. G. Human platelet size, shape and related functions in health and disease.Physiol. Rev. 62, 185–261 (1982).

    PubMed  CAS  Google Scholar 

  7. Johnson, P. C., Ware, J. A., Clivden, P. B., Smith, M., Pvorak, A. M. and Salzman, E. W., Measurement of ionized calcium in blood platelet with the photoprotein aequorin: comparison with quin 2.J. Biol. Chem. 260, 2069–2076 (1985).

    PubMed  CAS  Google Scholar 

  8. Katzenschlager, R., Weiss, K., Rogatti, W., Stelzeneder, M. and Sinzinger, H., Interaction between prostaglandin E1 and nitric oxide (NO).Thomb. Res. 62, 299–304 (1991).

    Article  CAS  Google Scholar 

  9. Kim, D. H., Ohnishi, T. and Ikemoto, N., Kinetic studies of calcium release from sarcoplasmic reticulumin vitro.J. Biol. Chem. 258, 9612–9668 (1983).

    Google Scholar 

  10. Ko, F. N., Sheu, S. J., Liu, Y. M., Huang, T. F. and Teng, C. M., Inhibition of rabbit platelet aggregation by 1,4-naphthoquinones.Thromb. Res. 57, 453–459 (1990).

    PubMed  Article  CAS  Google Scholar 

  11. Kovacs, T., Tordai, A., Szasz, I., Sarkadi, B. and Gardos, G., Membrane depolarization inhibits thrombin-induced calcium influx and aggregation in human platelets.FEBS Lett. 266, 171–174 (1989).

    Article  Google Scholar 

  12. Lasslo, A, and Quintana, R. P., Interaction dynamics of blood plateles with medicinal agents and other chemical entities. InBlood Platelet Function and Medicinal Chemistry (A. Lasslo, Ed.) Elsevier, New York, 1984, pp. 229–315.

    Google Scholar 

  13. Lind, C., Hochstein, P. and Ernster, L., DT-diaphorase as a quinone reductase: cellular control device against semiquinone and superoxide radical formation.Arch. Biochem. Biophys, 216, 178–105 (1982).

    PubMed  Article  CAS  Google Scholar 

  14. Mirabelli, F., Salis, A., Perotti, M., Taddei, F., Bellomo, G. and Orrenius, S., Alteration of surface morphology caused by the metabolism of menadione in mammalian cells are associated with the oxidation of critical sulfhydryl groups in cytoskeletal proteins.Biochem. Pharmacol. 37, 3423–3427 (1988).

    PubMed  Article  CAS  Google Scholar 

  15. Mirabelli, F., Salis, A., Vairetti, M., Bellomo, G., Thor, H. and Orrenius, S., Cytoskeletal alterations in human platelets exposed to oxidative stress are mediated by oxidative and Ca2+-dependent mechanisms.Arch. Biochem. Biophys. 270, 478–488 (1989).

    PubMed  Article  CAS  Google Scholar 

  16. Monks, T. J., Hanzlik, R. P., Cohen, G. M., Ross, K. and Graham, D. G., Contemporary issues in toxicology: Quinone chemistry and toxicity.Toxicol. Appl. Pharmacol. 112, 2–16 (1992).

    PubMed  Article  CAS  Google Scholar 

  17. Mustard, J. F., Perry, D. W., Ardile, N. G. and Packman, M. A., Preparation of suspension of washed platelets from humans.Br. J. Haematol. 22, 193–204 (1972).

    PubMed  Article  CAS  Google Scholar 

  18. Mustard, J. F. and Packman, M. A., Factors influencing platelet function: Adhesion release and aggregation.Pharmacol. Rev. 22, 97–187 (1979).

    Google Scholar 

  19. Nicotera, P., Bellomo, G. and Orrenius, S., Calcium-mediated mechanisms in chemically induced cell death.Annu. Rev. Pharmacol. Toxicol. 32, 449–470 (1992).

    PubMed  Article  CAS  Google Scholar 

  20. Puri, R. N., Zhon, F., Bradford, H. and Hu, C. H., Thrombin-induced platelet aggregation involves an indirect proteolytic cleavage by calpain.Arch. Biochem. Biophys. 271, 346–358 (1989).

    PubMed  Article  CAS  Google Scholar 

  21. Radomski, M. W., Palmer, R. M. J. and Moncada, S., Comparative pharmacology of endothelium-derived relaxing factor, nitric oxide and prostacyclin in platelets.Br. J. Pharmac. 92, 181–187 (1987).

    CAS  Google Scholar 

  22. Rink, T. J., Smith, S. W. and Tsien, R. Y., Cytoplasmic Free Ca+2 in human platelets: Ca+2 thresholds and Ca+2-independent activation for shape change and secretion.FEBS Lett. 148, 21–26 (1982).

    PubMed  Article  CAS  Google Scholar 

  23. Rink, T. J. and Sage, S. O., Calcium signaling in human platelets.Annu. Rev. Physiol. 52, 431–439 (1990).

    PubMed  Article  CAS  Google Scholar 

  24. Smith, P. F., Alberts, D. W. and Rush, G. F., Menadione-induced oxidative stress in hepatocytes isolated from fed and fasted rat: The role of NADPH-regenerating pathways.Toxicol. Appl. Pharmacol. 89, 190–201 (1987).

    PubMed  Article  CAS  Google Scholar 

  25. Stormorken, H., Platelets in hemostasis and thrombosis. InPlatelet Responses and Metabolism (Holmsen, H. Ed.), CRC Press, Boca Raton, 1984, pp. 3–32.

    Google Scholar 

  26. Thompson, A. R. and Harker, L. A. InManual of Hemostasis and Thrombosis. F. A. Davis Co., Philadelphia, 1987, pp. 57–85.

    Google Scholar 

  27. Thor, H., Smith, M.T., Hartzell, P., Bellomo, G., Jewell, S.A. and Orrenius, S., The metabolism of menadione (2-methyl-1,4-naphthoquinone) by isolated rat hepatocytes.J. Biol. Chem. 257, 12419–12425 (1982).

    PubMed  CAS  Google Scholar 

  28. Tsien, R. Y., Rozzan, T. and Rink, T. J., Calcium homeostasis in intact lymphocytes: Cytoplasmic free calcium monitored with a new intracellular trapped fluorescent indicator.J. Cell Biol. 94, 325–334 (1982).

    PubMed  Article  CAS  Google Scholar 

  29. Wefers, H. and Sies, H., Hepatic low-level chemiluminescence during redox cycling of menadione and the menadione-glutathione conjugate: Relation to glutathione and NAD(P)H quinone reductase (DT-diaphorase) activity.Arch. Biochem. Biophys. 224, 568–578 (1983).

    PubMed  Article  CAS  Google Scholar 

Download references

Author information

Affiliations

Authors

Corresponding author

Correspondence to Jin-Ho Chung.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Kim, KA., Kim, MJ., Ryu, CK. et al. Menadione-induced cytotoxicity in rat platelets: Absence of the detoxifying enzyme, quinone reductase. Arch. Pharm. Res. 18, 256–261 (1995). https://doi.org/10.1007/BF02976409

Download citation

Key words

  • Menadione
  • Platelets
  • Quinone reductase
  • Cytotoxicity