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Role of Reactive Oxygen Species and Glutathione in Inorganic Mercury-Induced Injury in Human Glioma Cells

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Abstract

The present study was undertaken to examine the role of reactive oxygen species (ROS) and glutathione (GSH) in glia cells using human glioma cell line A172 cells. HgCl2 caused the loss of cell viability in a dose-dependent manner. HgCl2-induced loss of cell viability was not affected by H2O2 scavengers catalase and pyruvate, a superoxide scavenger superoxide dismutase, a peroxynitrite scavenger uric acid, and an inhibitor of nitric oxide NG-nitro-arginine Methyl ester. HgCl2 did not cause changes in DCF fluorescence, an H2O2-sensitive fluorescent dye. The loss of cell viability was significantly prevented by the hydroxyl radical scavengers dimethylthiourea and thiourea, but it was not affected by antioxidants DPPD and Trlox. HgCl2-induced loss of cell viability was accompanied by a significant reduction in GSH content. The GSH depletion was almost completely prevented by thiols dithiothreitol and GSH, whereas the loss of viability was partially prevented by these agents. Incubation of cells with 0.2 mM buthionine sulfoximine for 24 hr, a selective inhibitor of γ-glutamylcysteine synthetase, resulted in 56% reduction in GSH content without any change in cell viability. HgCl2 resulted in 34% reduction in GSH content, which was accompanied by 59% loss of cell viability. These results suggest that HgCl2-induced cell death is not associated with generation of H2O2 and ROS-induced lipid peroxidation. In addition, these data suggest that the depletion of endogenous GSH itself may not play a critical role in the HgCl2-induced cytotoxicity in human glioma cells.

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REFERENCES

  1. Aschner, M. and Aschner, J. L. 1990. Mercury neurotoxicity: mechanisms of blood-brain barrier transport. Neurosci. Biobehav. Rev. 14:169–176.

    Google Scholar 

  2. Asano, S., Eto, K., Kurisaki, E., Gunji, H., Hiraiwa, K., Sato, M., Sato, H., Hasuike, M., Hagiwara, N., and Wakasa, H. 2000. Review article: acute inorganic mercury vapor inhalation poisoning. Pathol. Int. 50:169–174.

    Google Scholar 

  3. Olivieri, G., Brack, C., Muller-Spahn, F., Stahelin, H. B., Herrmann, M., Renard, P., Brockhaus, M., and Hock, C. 2000. Mercury induces cell cytotoxicity and oxidative stress and increases beta-amyloid secretion and tau phosphorylation in SHSY5Y neuroblastoma cells. J. Neurochem. 74:231–236.

    Google Scholar 

  4. Stacey, N. H. and Kappus, H. 1982. Cellular toxicity and lipid peroxidation in response to mercury. Toxicol. Appl. Pharmacol. 63:29–35.

    Google Scholar 

  5. LeBel, C. P., Ali, S. F., McKee, M., and Bondy, S. C. 1990. Organometal-induced increases in oxygen reactive species: the potential of 2′,7′-dichlorofluorescin diacetate as an index of neurotoxic damage. Toxicol. Appl. Pharmacol. 104:17–24.

    Google Scholar 

  6. Lund, B. O., Miller, D. M., and Woods, J. S. 1993. Studies on Hg(II)-induced H2O2 formation and oxidative stress in vivo and in vitro in rat kidney mitochondria. Biochem. Pharmacol. 45:2017–2024.

    Google Scholar 

  7. Hussain, S., Rodgers, D. A., Duhart, H. M., and Ali, S. F. 1997. Mercuric chloride-induced reactive oxygen species and its effect on antioxidant enzymes in different regions of rat brain. J. Environ. Sci. Health B 32:395–409.

    Google Scholar 

  8. Naganuma, A., Anderson, M. E., and Meister, A. 1990. Cellular glutathione as a determinant of sensitivity to mercuric chloride toxicity. Prevention of toxicity by giving glutathione monoester. Biochem. Pharmacol. 40:693–697.

    Google Scholar 

  9. Zalups, R. K. and Barfuss, D. W. 1995. Accumulation and handling of inorganic mercury in the kidney after coadministration with glutathione. J. Toxicol. Environ. Health 44:385–399.

    Google Scholar 

  10. Ballatori, N. and Clarkson, T. W. 1983. Biliary transport of glutathione and methylmercury. Am. J. Physiol. 244:G435–G441.

    Google Scholar 

  11. Refsvik, T. 1983. The mechanism of biliary excretion of methyl mercury: studies with methylthiols. Acta Pharmacol. Toxicol. 53:153–158.

    Google Scholar 

  12. Aihara, M. and Sharma, R. P. 1986. Effects of endogenous and exogenous thiols on the distribution of mercurial compounds in mouse tissues. Arch. Environ. Contam. Toxicol. 15:629–636.

    Google Scholar 

  13. Zalups, R. K. 2000. Molecular interactions with mercury in the kidney. Pharmacol. Rev. 52:113–143.

    Google Scholar 

  14. Berndt, W. O., Baggett, J. M., Blacker, A., and Houser, M. 1985. Renal glutathione and mercury uptake by kidney. Fundam. Appl. Toxicol. 5:832–839.

    Google Scholar 

  15. Baggett, J. M. and Berndt, W. O. 1986. The effect of depletion of nonprotein sulfhydryls by diethyl maleate plus buthionine sulfoximine on renal uptake of mercury in the rat. Toxicol. Appl. Pharmacol. 83:556–562.

    Google Scholar 

  16. Lash, L. H., Putt, D. A., and Zalups, R. K. 1999. Influence of exogenous thiols on inorganic mercury-induced injury in renal proximal and distal tubular cells from normal and uninephrectomized rats. J. Pharmacol. Exp. Ther. 291:492–502.

    Google Scholar 

  17. Philbert, M. A., Beiswanger, C. M., Waters, D. K., Reuhl, K. R., and Lowndes, H. E. 1991. Cellular and regional distribution of reduced glutathione in the nervous system of the rat: histochemical localization by mercury orange and o-phthaldialdehyde-induced histofluorescence. Toxicol. Appl. Pharmacol. 107:215–227.

    Google Scholar 

  18. Aschner, M. and LoPachin, R. M., Jr. 1993. Astrocytes: targets and mediators of chemical-induced CNS injury. J. Toxicol. Environ. Health 38:329–342.

    Google Scholar 

  19. Aschner, M. 1996. Astrocytes as modulators of mercuryinduced neurotoxicity. Neurotoxicology 17:663–669.

    Google Scholar 

  20. Denizot, F. and Lang R. 1986. Rapid colorimetric assay for cell growth and survival. Modifications to the tetrazolium dye procedure giving improved sensitivity and reliability. J. Immunol. Methods 89:271–277.

    Google Scholar 

  21. Bass, D. A., Parce, J. W., Dechatelet, L. R., Szejda, P., Seeds, M. C., and Thomas, M. 1983. Flow cytometric studies of oxidative product formation by neutrophils: a graded response to membrane stimulation. J. Immunol. 130:1910–1917.

    Google Scholar 

  22. Lebel, C. P., Ischiropoulos, H., and Bondy, S. C. 1992. Evaluation of the probe 2′,7′-dichlorofluorescin as an indicator of reactive oxygen species formation and oxidative stress. Chem. Res. Toxicol. 5:227–231.

    Google Scholar 

  23. Rosenkranz, A. R., Schmaldienst, S., Stuhlmeier, K. M., Chen, W., Knapp, W., and Zlabinger, G. J. 1992. A microplate assay for the detection of oxidative products using 2′,7′-dichlorofluorescin-diacetate. J. Immunol. Methods 156:39–45.

    Google Scholar 

  24. Hagar, H., Ueda, N., and Shah, S. V. 1996. Role of reactive oxygen metabolites in DNA damage and cell death in chemical hypoxic injury to LLC-PK1 cells. Am. J. Physiol. 271(1 Pt 2):F209–F215.

    Google Scholar 

  25. Lund, B. O., Miller, D. M., and Woods, J. S. 1991. Mercury- induced H2O2 production and lipid peroxidation in vitro in rat kidney mitochondria. Biochem. Pharmacol. 42:S181–S187.

    Google Scholar 

  26. Nath, K. A., Croatt, A. J., Likely, S., Behrens, T. W., and Warden, D. 1996. Renal oxidant injury and oxidant response induced by mercury. Kidney Int. 50:1032–1043.

    Google Scholar 

  27. Woolfson, R. G., Qasim, F. J., Thiru, S., Oliveira, D. B., Neild, G. H., and Mathieson, P. W. 1995. Nitric oxide contributes to tissue injury in mercuric chloride-induced autoimmunity. Biochem. Biophys. Res. Commun. 217:515–521.

    Google Scholar 

  28. Yamashita, T., Ando, Y., Sakashita, N., Hirayama, K., Tanaka, Y., Tashima, K., Uchino, M., and Ando, M. 1997. Role of nitric oxide in the cerebellar degeneration during methylmercury intoxication. Biochim. Biophys. Acta 1334:303–311.

    Google Scholar 

  29. Garcia-Ruiz, C., Colell, A., Mari, M., Morales, A., and Fernandez-Checa, J. C. 1997. Direct effect of ceramide on the mitochondrial electron transport chain leads to generation of reactive oxygen species. Role of mitochondrial glutathione. J. Biol. Chem. 272:11369–11377.

    Google Scholar 

  30. Kartha, V. N. and Krishnamurthy, S. 1978. Effect of vitamins, antioxidants and sulfhydryl compounds on in vitro rat brain lipid peroxidation. Int. J. Vitam. Nutr. Res. 48:38–43.

    Google Scholar 

  31. Robb, S. J. and Connor, J. R. 1998. An in vitro model for analysis of oxidative death in primary mouse astrocytes. Brain Res. 788:125–132.

    Google Scholar 

  32. Lin, A. M. and Ho, L. T. 2000. Melatonin suppresses ironinduced neurodegeneration in rat brain. Free Radic. Biol. Med. 28:904–911.

    Google Scholar 

  33. Murer, H. and Burckhardt, G. 1983. Membrane transport of anions across epithelia of mammalian small intestine and kidney proximal tubule. Rev. Physiol. Biochem. Pharmacol. 96:1–51.

    Google Scholar 

  34. Paller, M. S. 1985. Free radical scavengers in mercuric chloride-induced acute renal failure in the rat. J. Lab. Clin. Med. 105:459–463.

    Google Scholar 

  35. Halliwell, B., Gutteridge, J. M. C., and Cross, C. E. 1992. Free radicals, antioxidants, and human disease: Where are we now? J. Lab. Clin. Med. 119:598–620.

    Google Scholar 

  36. Min, S. K., Kim, S. Y., Kim, C. H., Woo, J. S., Jung, J. S., and Kim, Y. K. 2000. Role of lipid peroxidation and poly (ADPribose) polymerase activation in oxidant-induced membrane transport dysfunction in opposum kidney (OK) cells. Toxicol. Appl. Pharmacol. 166:196–202.

    Google Scholar 

  37. Kirkland, J. B. 1991. Lipid peroxidation, protein thiol oxidation and DNA damage in hydrogen peroxide-induced injury to endothelial cells: role of activation of poly(ADP-robose)polymerase. Biochim. Biophys. Acta 1092:319–325.

    Google Scholar 

  38. Thies, R. L. and Autor, A. P. 1991. Reactive oxygen injury to cultured pulmonary artery endothelial cells: mediation by poly(ADPribose) polymerase activation causing NAD depletion and altered energy balance. Arch. Biochem. Biophys. 286:353–363.

    Google Scholar 

  39. Yamamoto, K., Tsukidate, K., and Farber, J. L. 1993. Differing effects of the inhibition of poly(ADP-ribose) polymerase on the course of oxidative cell injury in hepatocytes and fibroblasts. Biochem. Pharmacol. 46:483–491.

    Google Scholar 

  40. Kappus, H. and Reinhold C. 1994. Heavy metal-induced cytotoxicity to cultured human epidermal keratinocytes and effects of antioxidants. Toxicol. Lett. 71:105–109.

    Google Scholar 

  41. Jones, M. M. and Basinger, M. A. 1980. Chelate therapy for type b metal ion poisoning. A. E. Martell (ed.). in Inorganic chemistry in biology and medicine. Washington, D.C.: American Chemical Society 335.

    Google Scholar 

  42. Vallee, B. L. and Ulmer, D. D. 1972. Biochemical effects of mercury, cadmium, and lead. Annu. Rev. Biochem. 41:91–128.

    Google Scholar 

  43. Gstraunthaler, G., Pfaller, W., and Kotanko, P. 1983. Glutathione depletion and in vitro lipid peroxidation in mercury or maleate induced acute renal failure. Biochem. Pharmacol. 32:2969–2972.

    Google Scholar 

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Authors and Affiliations

  1. Department of Neurosurgery, College of Medicine, Pusan National University, Pusan, 602-739, Korea

    Young Woo Lee

  2. Department of Physiology, College of Medicine, Pusan National University, Pusan, 602-739, Korea

    Mi Suk Ha & Yong Keun Kim

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Lee, Y.W., Ha, M.S. & Kim, Y.K. Role of Reactive Oxygen Species and Glutathione in Inorganic Mercury-Induced Injury in Human Glioma Cells. Neurochem Res 26, 1187–1193 (2001). https://doi.org/10.1023/A:1013955020515

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