Paraquat에 의한 사람 임파구 DNA 손상에 대한 환원전리수의 보호효과

Protective Effect of Electrolyzed Reduced Water on the Paraquat-induced Oxidative Damage of Human Lymphocyte DNA

  • 박은주 (경남대학교 식품영양학과) ;
  • 류근걸 (순천향대학교 신소재화학공학부) ;
  • 이윤배 (순천향대학교 신소재화학공학부) ;
  • 이종권 (순천향대학교 신소재화학공학부) ;
  • 이미영 (순천향대학교 생명과학부)
  • Park, Eun-Ju (Department of Food and Nutritional Sciences, Kyungnam University) ;
  • Ryoo, Kun-Kul (Division of Material & Chemical Engineering, Soonchunhyang University) ;
  • Lee, Yoon-Bae (Division of Material & Chemical Engineering, Soonchunhyang University) ;
  • Lee, Jong-Kwon (Division of Material & Chemical Engineering, Soonchunhyang University) ;
  • Lee, Mi-Young (Division of Life Science, Soonchunhyang University)
  • 발행 : 2005.06.30

초록

본 연구에서는 매우 낮은 음의 산화환원전위를 나타내는 환원전리수가 paraquat에 의한 사람 임파구 DNA의 손상에 미치는 영향을 Comet assay를 사용하여 조사하였다. 환원전리수 처리에 의해서 paraquat에 의해 산화적으로 손상된 DNA가 회복되는 정도를 손상된 DNA로 인한 꼬리부분의 형광광도의 %비율로 나타내었다. Comet assay는 개별 세포의 DNA의 산화적 손상을 측정하는데 널리 사용되고 있다. 사람 임파구에 다양한 농도의 paraquat을 $37^{\circ}C$에서 30분간 처리한 후, 환원전리수를 첨가하여 30분간 반응시켰다. 그 결과 paraquat에 의한 임파구 DNA의 손상은 paraquart 농도증가에 의존적으로 증가하였다. 그러나 환원전리수를 처리한 결과 DNA의 산화적 손상이 paraquat 미처리 대조군 수준으로 거의 다 복구되었다.

Electrolyzed reduced water (ERW), showing extremely negative oxidation-reduction potential, was used to investigate the effects of paraquat-induced damages on DNA from human lymphocyte. The effect of ERW on paraquat-induced oxidative DNA damage in human lymphocytes was evaluated by Comet assay (single-cell gel electrophoresis) quantified as percentage fluorescence in tail. Comet assay has been used widely to assess the level of the DNA damage in individual cells. Lymphocytes were oxidatively challenged with various concentrations of paraquat for 30 min at $37^{\circ}C$, and were then treated with electrolyzed reduced water for 30 min. The oxidative DNA damage by paraquat, as indicated by the fluorescent tail in DNA, increased in a dose-dependent manner. However, oxidative damage of the DNA was almost completely prevented upon treatment with electrolyzed reduced water.

키워드

참고문헌

  1. Lheureus, P., Leduc, D., Vanbinst, R. B. and Askenasi, R. (1995) Survival in a case of massive paraquat ingestion. Chest 107, 285-289 https://doi.org/10.1378/chest.107.1.285
  2. Lin, J. L., Leu, M. L., Liu, Y. C. and Chen, G. H. (1999) A prospective clinical trial of pulse therapy with glucocorticoid and cyclophosphamide in moderate to severe paraquat-poisoned patients. Am. J. Resp. Crit. Care 159, 357-360
  3. Bus, J. S., Aust, S. D. and Gibson, J. E. (1974) Superoxide and singlet oxygen catalyzed lipid peroxidationi as a possible mechanism for paraquat toxicity. Biochem. Biophys. Res. Commun. 58, 749-755 https://doi.org/10.1016/S0006-291X(74)80481-X
  4. Babior, B. M. and Peters, W. A. (1981) The $O^{2-}$ producing enzyme of human neutrophils: further properties. J. Biol. Chem. 256, 2321-2323
  5. Stocker, R. and Frei, B. (1991) In Oxidative stress: endogenous antioxidant defences in human blood plasma. Helmut, S. (ed.), San Diego, Academic Press INC, pp. 218-238
  6. Hong, S. Y., Yang, D. H. and Kim, Y. T. (1996) Successful management of severe paraquat poisoning with free radical scavenger. Korean J. Med. 51, 99-107
  7. Kang, S. A., Jang, Y. J. and Park, H. (1998) In vivo dual effects of vitamin C on paraquat-induced lung damage:dependence on released metals from the damaged tissue. Free Radical Res. 28, 93-107 https://doi.org/10.3109/10715769809097880
  8. Ryoo, K. K., Kang, B. and Sumida, S. (2002) Electrolyzed water as an alternative for environmentally-benign semiconductor cleaning. Mater. Res. Soc. 17, 1298-1304 https://doi.org/10.1557/JMR.2002.0194
  9. Huang, K. C., Yang, C. C., Lee, K. T. and Chien, C. T. (2003) Reduced hemodialysis-induced oxidative stress in end-stage renal disease patients by electrolyzed reduced water. Kidney Int. 64, 704-714 https://doi.org/10.1046/j.1523-1755.2003.00118.x
  10. Xin, H., Zheng, Y. J., Hajime, N. and Han, Z. G. (2003) Effect of electrolyzed water and hydrocolloid occlusive dressings on excised burn-wounds in rats. Chin. J. Traumatol. 6, 234-237
  11. Sanetaka, S., Shigeru, K., Mariko, N., Takumi, M., Kenichi, K., Miho, G., Hidemitsu, H., Kazumichi, O., Shinkatsu, M. and Yoshinori, K. (1997) Electrolyzed-reduced water scavenges active oxygen species and protects DNA from oxidative damage. Biochem. Biophys. Res. Commun. 234, 269-274 https://doi.org/10.1006/bbrc.1997.6622
  12. Park, E. J. and Kang, M. H. (2002) Application of the alkaline comet assay for detecting oxidative DNA damage in human biomonitoring. Korean J. Nutr. 35, 213-222
  13. Singh, P. N., McCoy, M. T., Tice, R. R. andchneider, E. L. (1988) A simple technique for quantitation of low levels of DNA damage in individual cells. Exp. Cell. Res. 175, 184-191 https://doi.org/10.1016/0014-4827(88)90265-0
  14. Cotelle, S. and Ferard, J. F. (1999) Comet assay in genetic ecotoxicology: a review. Environ. Mol. Mutagen. 34, 246-255 https://doi.org/10.1002/(SICI)1098-2280(1999)34:4<246::AID-EM4>3.0.CO;2-V
  15. Friedmann, B., Caplin, M., Hartley, J. A. and Hochhauser, D. (2004) Modulation of DNA repair in vitro after treatment with chemotherapeutic agents by the epidermal growth factor receptor inhibitor Gefitinib (ZD1839). Clin. Cancer Res. 10, 6476-6486 https://doi.org/10.1158/1078-0432.CCR-04-0586
  16. Chakraborty, S., Roy, M. and Bhattacharya, R. K. (2004) Prevention and repair of DNA damage by selected phytochemicals as measured by single cell gel electrophoresis. J. Environ. Pathol. Toxicol. Oncol. 23, 215-226 https://doi.org/10.1615/JEnvPathToxOncol.v23.i3.50
  17. Azqueta, A., Pachon, G., Cascante, M., Creppy, E. E. and de Cerain, A. L. (2005) DNA damage induced by quinoxaline 1,4-di-N-oxide derivative (hypoxic selective agent) in Caco-2-cells evaluated by the comet assay. Mutagenesis in press
  18. Kleinsasser, N. H., Wallner, B. C., Wagner, C., Kastenbauer, E. R. and Harreus, U. A. (2004) DNA repair capacity in lymphocyte of nasopharyngeal cancer patients. Eur. Arch Otorhinol. in press
  19. Ryoo, K. K., Lee, Y. B., Lee, J. K. and Lee, M. Y. (2005) Permeability and dissolvability of cathodic electrolyzed water for electrophoretic gel and green tea components. J. Korean Acad. Indus. Soc. 6, 87-93
  20. Lee, H. J., Kim, D. Y. and Chang, C. C. (1999) Antioxidant effects of Korean Red Ginseng components on the antioxidant enzymes activity and lipid peroxidation in the liver of mouse treated with paraquat. J. Ginseng Res. 23, 182-189
  21. Choi, J. H. and Oh, S. K. (1984) Studies on the anti-aging action of Korean Ginseng (II): The inhibitory effects of diol and triol saponins on lipoperoxide formation. Korean Biochem. J. 17, 445-455
  22. Frei, B., England, L. and Ames, B. N. (1989) Ascorbate is an outstanding antioxidant in human blood plasma. Proc. Natl. Acad. Sci. USA 86, 6377-6381
  23. Bateman, D. N. (1987) Pharmacological treatments of paraquat poisoning. Hum. Toxicol. 6, 57-62 https://doi.org/10.1177/096032718700600109
  24. Sittiount, C. (2005) Paraquat poisoning. Resp. Care 50, 383- 385
  25. Oho, J. H., Chung, S. P., Lim, H., Noh, S. H., Kim, H. Y., Kim, S. H., Lee, H. S. and Min, J. S. (2000) Effect of antioxidant, U-74389G, on paraquat-intoxicated rats. J. Korean Soc. Emerg. Med. 11, 431-436
  26. Fujimoto, Y., Nakatani, E., Horinouchi, M., Okamoto, K., Sakuma, S. and Fujita, T. (1989) Inhibition of paraquat accumulation in rabbit kidney cortex slices by ascorbic acid. Res. Commun. Chem. Pathol. Parmacol. 65, 245-248
  27. Hung, F., Switek, K. A., Zakharov, L. N., Fronczek, F. R., Slebodnick, C., Lam, M., Golen, J. A., Bryant, W. S., Mason, P. E., Rheingold, A. L., Ashorassani, M. and Gibson, H. W. (2005) Bis (m-phenylene)-32-crown-10-based cryptands, powerful hosts for paraquat derivatives. J. Org. Chem. 70. 3231-3241 https://doi.org/10.1021/jo050187i
  28. Huang, F., Zakharov, L. N., Rheingold, A. L., Ashraf- Khorassani, M. and Gibson, H. W. (2005) Synthesis of a symmetric cylindrical bis (crown ether) host and its complexation with paraquat. J. Org. Chem. 70, 809-813 https://doi.org/10.1021/jo048111u
  29. Ayaki, H., Lee, M. J., Sumino, K. and Nishio, H. (2005) Different cytoprotective effect of antioxidants and change in the iron regulatory system in rodent cells exposed to paraquat or formaldehyde. Toxicology 208, 73-79 https://doi.org/10.1016/j.tox.2004.11.008