생명과학회지 (Journal of Life Science)

  • 제16권2호
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  • Pages.259-265
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  • 2006
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  • 1225-9918(pISSN)
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  • 2287-3406(eISSN)
한국생명과학회 (Korean Society of Life Science)
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Streptozotocin 유도 당뇨성 흰쥐에서 methotrexate의 신독성 생성기전에 관한 연구

The Mechanism of Nephrotoxicity Formation of Methotrexate in STZ-Induced Hyperglycemic Rats

  • 발행 : 2006.04.01
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초록

엽산과 유사한 구조를 가져 관절염 치료제로 쓰이는 methotrexate (MTX)의 당뇨병 쥐에서의 신독성 기전을 구명할 목적으로 실험동물 정상군과 STZ로 당뇨를 유발한군에 MTX률 투여하여 신장 독성을 유발하여 혈중 생화학적 변화를 관찰하고 free radical의 생성계와 해독계의 활성에 미치는 영향을 관찰하였다. 혈중 신장기능 지표 효소 및 신장조직의 지질과산화 함량이 정상군에 MTX를 투여한 군에 비해 당뇨쥐에 MTX를 투여한 군에서 현저히 증가하였다. 당뇨쥐에 MTX를 투여함으로써 활성산소 생성계인 phase I 단계 중 cytosol 효소계 활성이 정상쥐에 MTX를 투여한 군에 비하여 현저히 증가하였고, phase II 단계 중 glutathione S-transferase의 활성이 정상쥐에 MTX를 투여하였을때에 비하여 당뇨쥐에 MTX를 투여하였을때 현저히 감소하였다. 신조직 중 glutathione의 함량 또한 당뇨쥐에 MTX를 투여했을때 현저히 감소하였고 이러한 결과는 $\gamma$-glutamylcysteine synthetase와 glutathione reductase의 활성을 감소시킨 결과로 생각된다. 이상의 결과를 종합하여 볼때 당뇨쥐에 MTX를 투여하면 신독성이 증가함을 확인할 수 있었고, 그 기전은 cytosol계 효소 활성을 증가시켜 MTX의 대사계를 촉진시킴으로 독성물질의 생성을 증가시키고, 이로 인한 신조직중의 지질과산화 함량의 증가는 glutathione의 함량 감소에 의해 나타나는 결과로 사료되어진다.

This study is investigated the effect on mechanism of nephrotoxicity formation of methotrexate(MTX) by hyperglycemic by streptozotocin(STZ). MTX was injected daily at two doses of 3, 6 mg/kg for 1 week in STZ-induced hyperglycemic rats. Activities of BUN, creatinine and LDH were significantly increased by treatment with MTX in STZ-induced diabetic group when compared to MTX treatment group in normal rats' Renal lipid peroxide content and activities of cytosolic enzyme were significantly increased in the treatment of MTX in diabetic group. The concentration of glutathione and glutathione biosynthesis enzymes were decreased by treatment with MTX in STZ-induced diabetic group. These results suggest that nephrotoxicity of MTX in STZ-induced hyperglycemic rat was caused by activation of renal metabolizing enzymes in cytosol and decrease of glutathione concentration.

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참고문헌

  1. Aebi, H. 1974. Catalase. In 'Methods of enzymetic analysis' Bergmeyer, H.U., Academic Press, New York., 2, 673-684
  2. Bidlack, W .R. and G. L. Lowry, 1982. Multiple drug metabolism: p-Nitroanisole reversal off acetone enhanced aniline hydroxylation. Biochemical Pharmacology 31, 311-317 https://doi.org/10.1016/0006-2952(82)90176-9
  3. Cabaud, P. G. and F. Wroblewski. 1958. The determination of serum lactate dehydrogenase. American Journal Clinical Pathology 30, 234-236 https://doi.org/10.1093/ajcp/30.3.234
  4. Chabner, B. A., F. Mayers and C. N. Coleman. 1975. Antineoplasticagents. New England Journal Medicin 292, 1107-1113 https://doi.org/10.1056/NEJM197505222922107
  5. Chaney, A. L. and E. P. Marbach. 1962. Modified reagents for determination of urea and ammonia, Clinica Chimica Acta 8, 130-132
  6. Charles, E. M. and G. L. Robert. 1962. Hepatic glutathione reductase. Journal of Biological Chemistry 237, 1589-1595
  7. Ellman, G. L. 1959. Tissue sulfhydryl group. Archives of Biochemistry and Biophysics 80, 70-77
  8. Esterbauer, H., R. J. Schaur and H. Zuller. 1991. Chemistry and biochemistry of 4-hydroxynonenal, malondialdehyde and related aldehyde. Free Radical Biology of Medicine 11, 81-128 https://doi.org/10.1016/0891-5849(91)90192-6
  9. Garcia, M. J., P. M. Mcnamara, T. Gordon and W. B. Kannel. 1974. Morbidity in mortality in diabetes in the framigham population. Diabetes 23, 105-108 https://doi.org/10.2337/diab.23.2.105
  10. Habig, W. H., M. J. Pabist and W. B. Jakoby. 1974. Glutathione S-transferase, The first enzymatic step in mercapturic acid formation., Journal Biological Chemistry 249, 7130-7139
  11. Ji, S., Lemaster, J. J., V. Christenson. and R. Thurman, 1990. Periportal and pericental pyrimidine nucleotide fluorescence from the surface of the perfused liver. Evaluation of the hypothesis that chronic treatment with ethanol produces pericentral hypoxia. Proceedings of the National Academy of Science U.S.A. 79, 5415-5419
  12. Kannel, W. B. and D. L. Mcgee. 1979. Diabetes and cardio vascular disease. J.A. H.A. 241, 2053-2038
  13. Kinney, C. S. and L. M. Morse. 1964. Effect of a folic acid antagonist in rat. Journal of Nutrition 84, 288-294 https://doi.org/10.1093/jn/84.3.288
  14. Lowry, O. H., R. R. Nitra, M .L. Wu, W. S. Hixon and E. J. Crawford. 1954. The guantitative histochemistry of brain, enzyme measurements. Journal Biological Chemistry 207, 19-37
  15. Lane, R. H., R. J. Ramirez, A. E. Tsirka, J. L. Kloesz, M. K. Malaughlin. E. M. Gruetzmacher and S. U. Devaskar. 2001. Uterplacental insufficiency lowers the threshold towerds hypoxia-induced cerebral apoptosis in growth-related fetal rats. Brain Reserch 895, 186-193 https://doi.org/10.1016/S0006-8993(01)02074-1
  16. Lawrene, R. A. and R. F. Burk. 1976. Glutathione peroxidase activity in selenium deficiet rat liver. Biochemical and Biophysical Research Communications 71, 952-958 https://doi.org/10.1016/0006-291X(76)90747-6
  17. Lee, S. I. 2004. Diabetes and the secondary complication, Korea Visuals, pp. 141-148
  18. Little, C. and P. J. O'Brine. 1968. An intracellular GSH -peroxidase with lipid peroxide substrate. Biochemical and Biophysical Research Communications 31, 145-150 https://doi.org/10.1016/0006-291X(68)90721-3
  19. Lowry, O. H., N. J., Rosebrough, A. L. Farr and Rendall, R. J. 1951. Protein Measurement with folin phenol reagent. Journal Biological Chemistry 193, 265-275
  20. Marklund, S. and G. Marklund. 1974. Involvement of the superoxide anion radical in the autooxidation of pyrogallol and a convenien assay for superoxide dismutase. European Journal Biochemistry 47, 469-474 https://doi.org/10.1111/j.1432-1033.1974.tb03714.x
  21. McCord, J. M. 1985. Oxygen derived free radicals in postischemic tissue injury. New England Journal Medicine 312, 159-163 https://doi.org/10.1056/NEJM198501173120305
  22. Meister, A. and P. G. Richman. 1975. Regulation of ${\gamma}-glutamylcysteine$ synt-hesis by nonallosteric feedback inhi- bition by glutathione. Journal of Biological Chemistry 250, 1422-1426
  23. Meister, A. and M. E. Anderson. 1983. Glutathione. Annual Review of Biochemistry 52, 711-760 https://doi.org/10.1146/annurev.bi.52.070183.003431
  24. Mielants, H., E .M., Veys, C. Straeten C. Ackerman .and S. Goemaere. 1991. The efficacy and toxicity of constant low dose of methotrexate as a treatment for intractable rheumatoid arthritis. The Journal of Rheumatology 18, 978-983
  25. Nash, T. 1953. The colorimetric estimation of formaldehyde by means of the hentisch reaction. Journal of Biological Chemistry 55, 416-421
  26. Ohkawa, H., N. Ohishi and K. Yaki. 1979 Assay for lipid peroxide in animal tissues by thiobarbituric acid reaction. Analytical Biochemistry 95, 351-358 https://doi.org/10.1016/0003-2697(79)90738-3
  27. Oyanagui, Y. 1984. Reevaluation of assay methods and establishment of kit for superoxide dismutase activity. Analytical Biochemistry 142, 290-296 https://doi.org/10.1016/0003-2697(84)90467-6
  28. Paglia, E .D. and W. N. Valentine. 1967. Studies on the quantitative and qualitayive charaterization of erythro- cytes glutathione peroxidase. Journal of Laboratory and Clinical Medicine 70, 158-169
  29. Rajagopalan, K. V., I Fridovich and P. Handler. 1962. Hepatic aldehyde ox-idase. In Purificatio and properties, Journal of Biological Chemistry 237, 922-928
  30. Reitman, S. and S. K. Frankel. 1957. A colorimetric method for determination of serum glutamic oxaloacetic and glutamic pyruvic trasaminase. American Journal of Clinical Pathology 28, 58-63
  31. Salaffi, F., M. Carotti, A. Sartini and C. Cervini. 1995. A prospective study of the long-term efficacy and toxicity of low-dose methotrexate in rheumatoid arthritis. Clinical and Experimental Rheumatology 13, 23-38
  32. Slot, C. 1973. Determination of serum creatinine by a direct colorimetric method. Clinica Chimica Acta 43, 305-310 https://doi.org/10.1016/0009-8981(73)90466-X
  33. Stephen, B. and K. Rita. 1980. Influence on methotrexate pharmacokinetics. European Journal of Cancer 16, 1427-1432 https://doi.org/10.1016/0014-2964(80)90051-1
  34. Stirpe, F. and C. E. Della. 1969. The regulation of rat liver xanthine oxidase Conversion in vitro of the enzyme activity from dehydrogenase(Type D) to oxidase(Type O). Journal of Biological Chemistry 244, 3855-3863
  35. Szasa, F. 1969. A Kinetic photometric method for serum glutamyltransferase. Clinical Chemistry 15, 124-136
  36. Weinblatt, M. E., B. N. Weissman D. E., Holdsworth, P. A. Fraser, A. L. Maier, K. R. Falchuk. and J. S. Coblyn. 1992. Long-term prospective study of methotrexate in the treatment of rheumatoid arthritis. Arthritis Rheumatism 35, 129-137 https://doi.org/10.1002/art.1780350202
  37. Wendel, A. and S. Feuerstein. 1981. Drug-induced lipid peroxidation in mice. Modulation by monooxygenase activity, glutathone and selenium status. Biochemical Pharmacology 30, 2513-2520 https://doi.org/10.1016/0006-2952(81)90576-1