지각 추출물이 quinone reductase 및 glutathion s-transferase의 유도활성에 미치는 영향

Induction of Quinone Reductase and Glutathion S-transferase in Hepatoma Cells by Citrus aurantium Linn (Jikak) Produced in Cheju Island

  • 유미희 (계명대학교 식품가공학 전공) ;
  • 이인선 (계명대학교 전통미생물자원개발 및 산업화연구(TMR) 센터)
  • Yu, Mi-Hee (Department of Food Science and Technology, School of Natural Science, Keimyung University) ;
  • Lee, In-Seon (The Center for Traditional Microorganism Resources, Keimyung University)
  • 발행 : 2005.04.30

초록

Phase II 효소계는 자연계에 존재하는 다양한 화학물질과 천연소재들에 의해 유도되며, 이들의 유도는 화학적 발암물질과 그 밖의 여러 가지 독성물질들로부터 생체를 보호하는데 중요한 역할을 한다. 특히, phase II 효소 중 QR은 quinone류 자체에 대한 보호효과가 있고 다른 암예방 효소계와 공통으로 유도되며, 항암 작용이 있는 많은 화합물에 의해 유도되어지기 때문에 암예방 물질 탐색의 지표가 되는 대표적인 효소로 많이 이용된다. 본 연구에서는 제주도에서 자생하는 귤류 중 지각을 이용하여 메탄올추출물을 제조한 후 대표적인 암예방계 효소로 알려진 quinone reductase 및 glutathion S-transferase의 유도 활성을 조사하였다. 우선 QR 및 GST 유도활성 측정에 앞서 각 시료의 Hepa 1c1c7 세포에 대한 독성을 조사하여 시료자체의 세포독성을 관찰하였으며, 이 결과를 바탕으로 QR, GST 유도 활성을 측정할 시 세포독성을 거의 나타내지 않는 각각 시료의 최대 처리 농도를 결정하였다. 총 6개의 지각 분획물 중 mouse 유래 Hepa 1c1c7 세포주에서는 hexane 충과 chloroform 충에서 QR, GST의 활성이 $200{\mu}g/mL$에서 각각 1.8배, 1.5배 이상으로 증가되어 높은 QR 및 GST의 유도활성을 보였다. 따라서 지각의 hexane 층이나 chloroform층에 존재하는 높은 QR및 GST의 유도활성이 나타나는 물질을 분리 정제하여 앞으로 계속적인 연구가 진행되어야 할 것이다.

참고문헌

  1. Kang GI. The conception by drugs of medical action and activity. pp. 245, 286-302. Hiseongchulpansa, Seoul, Korea (1993)
  2. Talalay P, Benson AM. Elvation of quinone reductase activity by anticarcinogenic antioxidants. Adv. Enzyme Reg. 20: 287-300 (1982) https://doi.org/10.1016/0065-2571(82)90021-8
  3. Wefers H, Komai T, Talalay P, Sies H. Protection against reactive oxygen species by NAD(P)H: quinone recuctase induced by the dietary antioxidant butylated hydroxyanisole (BHA). FEBS Lett. 169: 63-66 (1984) https://doi.org/10.1016/0014-5793(84)80290-2
  4. Prochaska HJ, Santamaria AB. Direct measurement of NAD(P)H: Quinone reductase from cells cultured in microtiter well: A screening assay for anticarcinogenic enzyme inducers. Anal. Biochem. 169: 328-336 (1988) https://doi.org/10.1016/0003-2697(88)90292-8
  5. Zhang Y, Talalay P, Cho CG, Posner GH. A major inducer of anticarcinogenic protective enzymes from broccoli: Islation and elucidation of structure. Proc. Natl. Acad. Sci. USA 89: 2399-2403 (1992)
  6. Prochaska HJ, Talalay P. Regulatory mechanisms of monofunctional and bifunctional anticarcinogenic enzyme inducers in murine liver. Cancer Res. 48: 4776-4782 (1988)
  7. Kwak YJ, Jun HJ, Kwon TW, Kim JS. Modulation of anticarcinogenic enzyme and plasma testosterone level in male mouse fed leek-supplemented diet. Korean J. Food Sci. Nutr. 27: 968-972 (1998)
  8. Mehta RG, Liu J, Constantinou A, Thomas CF, Hawthorne M, Gerhuser C, Pezzuto, JM, Moon RC, Moriarty RM. Cancer chemopreventive activity of brassinin, a phytoalexin from cabbage. Carcinogenesis 16: 399-404 (1995) https://doi.org/10.1093/carcin/16.2.399
  9. Gerhauser C, You M, Liu J, Moriarty RM, Hawthorne M, Mehta RG, Moon RC, Pezzuto JM. Cancer chemopreventive potential of sulforamate, a novel analogue of sulforaphane that induces phase 2 drug-metabolizing enzymes. Cancer Res. 57: 272-278 (1997)
  10. Kim SM, Ryu SH, Choi RD, Kim SS, Kim JH, Kim JS. Screening for Korean vegetables with anticarcinogenic enzyme inducing activity using cell culture system. Korean J. Food Sci. Nutr. 27: 277-28l (1998)
  11. Koh JS, Kim SH. Physicochemical properties and chemical compositions of citrus fruits produced in Cheju, Agric. Chem. Biotechnol. 38: 541-545 (1995)
  12. Hwang HJ, Yoon KR. Carotenoid pigment of citrus fruits cultivated in Korea. Food Biotechnol. 27: 950-957 (1995)
  13. Eun JB, Jung YM, Woo GJ. Identification and determination of dietary fibers and flavonoids and peel of Korean Tangerine (Citrus aurantium var.). Korean J. Food Sci. Technol. 28: 371-377 (1996)
  14. Oshiba J, Kato M. Nutritional regulation III, depression of cardiac action by the isolated naringin, Mukogawa Joshidaigaku Kiyo. 29: 1-8 (1981)
  15. Oh HS, Park WB, An YS, Oh MC, Oh CK, Kim SH. Antimicrobial activity of extracts from citrus seeds. Korean J. Culinary Res. 9: 69-80 (2003)
  16. Green LM, Reade JL, Ware CF. Rapid colormetric assay for cell viability: Application to the quantitation of cytotoxic and growth inhibitory lympokines. J. Immunol. Method 70: 257-268 (1984) https://doi.org/10.1016/0022-1759(84)90190-X
  17. Benson A, Hunkeler MJ, Talay P. Increase of NAD(P)H: qunone reductase by dietary antioxidants. Possible role in protection against carcinogenesis and toxicity. Proc. Natl. Acad. Sci. USA 77: 5216-5220 (1980)
  18. Habig WH, Pabist MJ, Iakoby WB. Glutathione S-transferase: The first enzymetic step in merapturic acid formation. J. BioI. Chem. 25: 7130-7139 (1974)
  19. Drysdale BE, Zacharchuk CM, Okajima M, Shin HS. In Methods in Enaymology (Di Sabato G, and Eberse T., Eds.). Academic press: Orlando. FL. 132: 549-555 (1986)
  20. Kim JS, Nam YJ, Kwon TW. Induction of quinone reductase activity by genistein, soybean isoflavone. Food Sci. Biotech. 5: 70-75 (1996)
  21. Kim HS, Kacew S, Lee BM. In vitro chemopreventive effects of plant polysaccharides (Aloe barbadensis Miller, Lentinus edodes, Ganoderma lucidum and Corious versicolor). Carcinogenesis 20: 1637-1640 (1999) https://doi.org/10.1093/carcin/20.8.1637
  22. Williamson G, DuPont MS, Wanigatunga S, Heaney RK, Musk SRR, Fenwick GR, Hodes MJC. Induction of glutathione S-transferase activity in hepG2 cells by extracts from fruits and vegetables. Food Chem. 60: 157-160 (1997) https://doi.org/10.1016/S0308-8146(95)00255-3
  23. Hashimoto K, Kawamata S, Usui N, Tanaka A, Uda Y. In vitro induction of the anticarcinogenic marker enzyme, quinone reductase, in human hepatoma cells by food extracts. Cancer Lett. 180: 1-5 (2002) https://doi.org/10.1016/S0304-3835(02)00018-6
  24. Talalay P, De Long MJ, Prochaska HJ. Identification of a common chemical signal regulating the induction of enzymes that protect against chemical carcinogenesis. Proc. Natl. Acad. Sci. USA 85: 8261-8265 (1988)
  25. Spencer SR, Wilczak CA, Talalay P. Induction of glutathione transferases and NAD(P)H: Quinone reductase by fumaric acid derivatives in rodent cells and tissues. Cancer Res. 50: 7871-7875 (1990)