Abstract
Intracellular oxidation and reduction (redox state) correspond closely to the surrounding environment. Most environmental factors affecting this balances such as oxidants, ultraviolet light, radioactive emissions, infections, and allergic reactions represent oxidative stress upon cells. We examined intracellular redox state after oxidative stress upon cultured human airway epithelial cells (Calu-3) by measuring reduced (GSH) or oxidized (GSSG) glutathione. We studied cytokine production, which is related to glutathione redox regulation, in response to ozone and also evaluated the effect of pretreatment with an ethyl ester of reduced glutathione (GSH-OEt) on cytokines. Ozone exposure (3.0 ppm, 3 min) time-dependently changed the redox state, while increasing production of interleukin(IL)-8 and IL-6, mRNA and protein. Treatment with GSH-OEt before ozone suppressed IL-8, but stimulated IL-6 production. Thus, oxidative stress affects intracellular glutathione redox state, in airway epithelial cells, activating signals to increase production of cytokine, modulation that may exacerbate respiratory symptoms.
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Epperlein, M. M., J. Nourooz-Zadeh, A. A. Noronha-Dutra, and N. Woolf. 1996. Nitric oxide in cigarette smoke as a mediator of oxidative damage. Int. J. Exp. Pathol. 77:197-200.
Repine, J. E., A. Bast, and I. Lankhorst. 1997. Oxidative stress in chronic obstructive pulmonary disease. Oxidative Stress Study Group. Am. J. Respir. Crit. Care Med. 156:341-357.
Barnes, P. J. 1995. Air pollution and asthma: Molecular mechanisms. Mol. Med. Today 1:149-155.
Bowler R. P. and J. D. Crapo. 2002. Oxidative stress in allergic respiratory diseases. J. Allergy Clin. Immunol. 110:349-356.
Nakamura, H., K. Nakamura, and J. Yodoi. 1997. Redox regulation of cellular activation. Annu. Rev. Immunol. 15:351-369.
Deneke, S. M. and B. L. Fanburg. 1989. Regulation of cellular glutathione. Am. J. Physiol. 257:L163-L173.
Meister, A. 1988. Glutathione metabolism and its selective modification. J. Biol. Chem. 263:17205-17208.
Meyer, M., H. L. Pahl, and P. A. Baeuerle. 1994. Regulation of the transcription factors NF-kappa B and AP-1 by redox changes. Chem. Biol. Interact. 91:91-100.
Rouse, J., P. Cohen, S. Trigon, M. Morange, A. Alonso-Llamazares, D. Zamanillo, T. Hunt, and A. R. Nebreda. 1994. A novel kinase cascade triggered by stress and heat shock that stimulates MAPKAP kinase-2 and phosphorylation of the small heat shock proteins. Cell 78:1027-1037.
Dobashi, K., M. Aihara, T. Araki, Y. Shimizu, M. Utsugi, K. Iizuka, Y. Murata, J. Hamuro, T. Nakazawa, and M. Mori. 2001. Regulation of LPS induced IL-12 production by IFN-gamma and IL-4 through intracellular glutathione status in human alveolar macrophages. Clin. Exp. Immunol. 124:290-296.
Utsugi, M., K. Dobashi, Y. Koga, Y. Shimizu, T. Ishizuka, K. Iizuka, J. Hamuro, T. Nakazawa, and M. Mori. 2002. Glutathione redox regulates lipopolysaccharide-induced IL-12 production through p38 mitogen-activated protein kinase activation in human monocytes: Role of glutathione redox in IFN-gamma priming of IL-12 production. J. Leukoc. Biol. 71:339-347.
Holtzman, M. J., L. M. Fabbri, B. E. Skoogh, P. M. O'Byrne, E. H. Walters, H. Aizawa, and J. A. Nadel. 1983. Time course of airway hyerresponsiveness induced by ozone in dogs. J. Appl. Physiol. 55:1232-1236.
Blomberg, A., I. S. Mudway, C. Nordenhall, H. Hedenstrom, F. J. Kelly, A. J. Frew, S. T. Holgate, and T. Sandstrom. 1999. Ozone-induced lung function decrements do not correlate with early airway inflammatory or antioxidant responses. Eur. Respir. J. 13:1418-1428.
Blomberg, A. 2000. Airway inflammatory and antioxidant responses to oxidative and particulate air pollutants—experimental exposure studies in humans. Clin. Exp. Allergy 30:310-317.
Mochizuki, H., Y. Ohki, H. Arakawa, K. Tokuyama, and A. Morikawa. 1999. Effect of ultrasonically nebulized distilled water on airway epithelial cell swelling in guinea pigs. J. Appl. Physiol. 86:1505-1512.
Mochizuki, H., Y. Ohki, H. Arakawa, M. Kato, K. Tokuyama, and A. Morikawa. 2002. Effect of inhaled indomethacin on distilled water-induced airway epithelial cell swelling. J. Appl. Physiol. 92:155-161.
Shen, B. Q., W. E. Finkbeiner, J. J. Wine, R. J. Mrsny, and J. H. Widdicombe. 1994. Calu-3: A human airway epithelial cell line that shows cAMP-dependent Cl-secretion. Am. J. Physiol. 266:L493-L501.
Buchmuller-Rouiller, Y., S. B. Corrandin, J. Smith, P. Schneider, A. Ransijn, C. V. Jongeneel, and J. Mauel. 1995. Role of glutathione in macrophage activation: Effect of cellular glutathione depletion on nitrite production and leishmanicidal activity. Cell Immunol. 164:73-80.
Tietze, F. 1969. Enzymic method for quantitative determination of nanogram amounts of total and oxidized glutathione: Applications to mammalian blood and other tissues. Anal. Biochem. 27:502-522.
Griffith, O. W. 1980. Determination of glutathione and glutathione disulfide using glutathione reductase and 2-vinylpyridine. Anal. Biochem. 106:207-212.
O'Mahony, J. and C. Hill. 2002. A real-time PCR assay for the detection and quantitation of Mycobacterium avium subsp. paratuberculosis using SYBR Green and the Light Cycler. J. Microbiol. Methods 51:283-293.
Koenig, M., K. S. Reynolds, W. Aldous, and M. Hickman. 2001. Comparison of Light-Cycler PCR, enzyme immunoassay, and tissue culture for detection of herpes simplex virus. Diagn. Microbiol. Infect. Dis. 40:107-110.
Murlas, C. G., Z. Lang, and G. J. Williams. 1992. Aerosolized neutral endoetidase reverses ozone-induced airway hyperreactivity to substance P. J. Appl. Physiol. 72:1133-1141.
Tarkington, B. K., T. R. Duvall, 1994. Last. Ozone exposure of cultured cells and tissues. Methods Enzymol. 234:257-265.
Lawrence, B. P., Y. Will, D. J. Reed, and N. I. Kerkvliet. 2000. Gamma-glutamyl transpeptidase knockout mice as a model for understanding the consequences of diminished glutathione on T-cell-dependent immune responses. Eur. J. Immunol. 30:1902-1910.
Suthanthiran, M., M. E. Anderson, V. K. Sharma, and A. Meister. 1990. Glutathione regulates activation-dependent DNA synthesis in highly purified normal human T lymphocytes stimulated via the CD2 and CD3 antigens. Proc. Natl. Acad. Sci. USA. 87:3343-3347.
Gmunder, H. and W. Droge. 1991. Differential effects of glutathione depletion on T-cell subsets. Cell Immunol. 138:229-237.
Ziegler, D. M. 1985. Role of reversible oxidation-reduction of enzyme thiols-disulfides in metabolic regulation. Annu. Rev. Biochem. 54:305-329.
Calhoun, W. J., H. E. Reed, D. R. Moest, and C. A. Stevens. 1992. Enhanced superoxide production by alveolar macrophages and air-space cells, airway inflammation, and alveolar macrophage density changes after segmental antigen bronchoprovocation in allergic subjects. Am. Rev. Respir. Dis. 145:317-325.
Thomas, J. A., B. Poland, and R. Honzatko. 1995. Protein sulfhydryls and their role in the antioxidant function of protein S-thiolation. Arch. Biochem. Biophys. 319:1-9.
Kunkel, S. L., T. Standiford, K. Kasahara, and R. M. Strieter. 1991. Interleukin-8 (IL-8): The major neutrophil chemotactic factor in the lung. Exp. Lung. Res. 17:17-23.
DeForge, L. E., J. C. Fantone, J. S. Kenney, and D. G. Remick. 1992. Oxygen radical scavengers selectively inhibit interleukin-8 production in human whole blood. J. Clin. Invest. 90:2123-2129.
Simeonova, P. P., and M. I. Luster. 1996. Asbestos induction of nuclear transcription factors and interleukin-8 gene regulation. Am. J. Respir. Cell. Mol. Biol. 15:787-795.
Simeonova, P. P., W. Toriumi, C. Kommineni, M. Erkan, A. E. Munson, W. N. Rom, and M. I. Luster. 1997. Molecular regulation of IL-6 activation by asbestos in lung epithelial cells: Role of reactive oxygen species. J. Immunol. 159:3921-3928.
Han, J., J. D. Lee, L. Bibbs, and R. J. Ulevitch. 1994. A MAP kinase targeted by endotoxin and hyperosmolarity in mammalian cells. Science 265:808-811.
Raingeaud, J., S. Gupta, J. S. Rogers, M. Dickens, J. Han, R. J. Ulevitch, and R. J. Davis. 1995. Pro-inflammatory cytokines and environmental stress cause p38 mitogen-activated protein kinase activation by dual phosphorylation on tyrosine and threonine. J. Biol. Chem. 270:7420-7426.
Hashimoto, S., K. Matsumoto, Y. Gon, T. Nakayama, I. Takeshita, and T. Horie. 1999. Hyperosmolarity-induced interleukin-8 expression in human bronchial epithelial cells through p38 mitogen-activated protein kinase. Am. J. Respir. Crit. Care Med. 159:634-640.
Schultheis, A. H., and D. J. Bassett. 1994. Guinea pig lung inflammatory cell changes following acute ozone exposure. Lung. 172:169-181.
Kelly, F. J., I. Mudway, A. Blomberg, A. Frew, and T. Sandstrom. 1999. Altered lung antioxidant status in patients with mild asthma. Lancet. 354:482-483.
Simeonova, P. P., S. Leonard, L. Flood, X. Shi, and M. I. Luster. 1999. Redox-dependent regulation of interleukin-8 by tumor necrosis factor-alpha in lung epithelial cells. Lab. Invest. 79:1027-1037.
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Todokoro, M., Mochizuki, H., Tokuyama, K. et al. Effect of Ozone Exposure on Intracellular Glutathione Redox State in Cultured Human Airway Epithelial Cells. Inflammation 28, 105–114 (2004). https://doi.org/10.1023/B:IFLA.0000033026.91221.ed
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DOI: https://doi.org/10.1023/B:IFLA.0000033026.91221.ed