Study of Skin Depigmenting Mechanism of the Ethanol Extract of Fagopyrum esculentum

교맥 에탄올 추출물의 피부 미백기전 연구

  • No, Seong-Taek (Department of Herbal Resources, Professional Graduate School of Oriental Medicine, Wonkwang University) ;
  • Kim, Dae-Sung (Department of Herbal Resources, Professional Graduate School of Oriental Medicine, Wonkwang University) ;
  • Lee, Seong-Jin (Department of the Third Medicine, Wonkwang University) ;
  • Park, Dae-Jung (Department of Herhology & Precriptionology, College of Oriental Medicine, Medicine, Sangji University) ;
  • Lee, Jang-Cheon (Department of Herhology & Precriptionology, College of Oriental Medicine, Medicine, Sangji University) ;
  • Lim, Kyu-Sang (Department of Herbal Resources, Professional Graduate School of Oriental Medicine, Wonkwang University) ;
  • Woo, Won-Hong (Department of Herbal Resources, Professional Graduate School of Oriental Medicine, Wonkwang University) ;
  • Mun, Yeun-Ja (Department of Herbal Resources, Professional Graduate School of Oriental Medicine, Wonkwang University)
  • 노성택 (원광대학교 한의학전문대학원 한약자원개발학과) ;
  • 김대성 (원광대학교 한의학전문대학원 한약자원개발학과) ;
  • 이성진 (원광대학교 한의학전문대학원 제3의학과) ;
  • 박대중 (상지대학교 한의과대학 본초방제학교실) ;
  • 이장천 (상지대학교 한의과대학 본초방제학교실) ;
  • 임규상 (원광대학교 한의학전문대학원 한약자원개발학과) ;
  • 우원홍 (원광대학교 한의학전문대학원 한약자원개발학과) ;
  • 문연자 (원광대학교 한의학전문대학원 한약자원개발학과)
  • Published : 2007.10.25

Abstract

The aim of this study was to investigate the effect of ethanol extract of Fagopyrum esculentum on the melanogenesis. To determine whether ethanol extract of Fagopyrum esculentum suppress melanin synthesis in cellular level, B16F10 melanoma cells were cultured in the presence of different concentrations of Fagopyrum esculentum ethanol extract. In the present study, we examined the effects of Fagopyrum esculentum ethanol extract on cell proliferation, melanin contents, tyrosinase activity, expression of melanogenic enzyme proteins including tyrosinase, tyrosinase-related protein 1 (TRP-1) and tyrosinase-related protein 2 (TRP-2). Cell proliferation was slightly increased by treatment with ethanol extract of Fagopyrum esculentum $(25-200 {\mu}g/m{\ell}).$ The ethanol extract of Fagopyrum esculentum effectively suppressed melanin contents at a dose of $100 {\mu}g/m{\ell}).$ It was observed that the color of cell pellets was totally whitened compared with the control. The ethanol extract of Fagopyrum esculentum inhibited tyrosinase activity, regulate melanin biosynthesis as the key enzyme in melanogenesis. Using western blot analysis, the ethanol extract of Fagopyrum esculentum dose-dependently decreased tyrosinase and TRP-1 protein levels, and tyrosinase and TRP-1 were detected in similar manner. ${\alpha}-MSH$ leads to a stimulation of melanin synthesis through increase of tyrosinase activity, melanin contents and cytoplasmic dendricity. In this study, ethanol extract of Fagopyrum esculentum down-regulated the ${\alpha}-MSH$-induced tyrosinase activity, melanin contents and cytoplasmic dendricity. Regarding protein levels of the melanogenic enzymes, the amounts of tyrosinase and TRP-1 was increased after incubation with a-MSH. The treatment of ethanol extract of Fagopyrum esculentum decreased the ${\alpha}-MSH$-induced expression levels of tyrosinase and TRP-1. These results suggest that the ethanol extract of Fagopyrum esculentum exerts its depigmenting effects through the suppression of tyrosinase, TRP-1 and cytoplasmic dendricity. And it may be a potent depigmetation agent in hyperpigmentation condition.

Keywords

References

  1. Lee, J.S., Ra, K.S., Son, H. Extraction and component sugar analysis of polysaccharides from buckwgeat. Korean J. Food Sci. Technol. 27: 860-865, 1995
  2. Lee, G.D., Yun, S.L., Kim, J.O., Heo, S.S., Seo, G.I. Monitoring on the tea with steaming and drying process of germinated byck wheat. J. korean Soc. Food Sci. Nutr. 33: 212-217, 2004 https://doi.org/10.3746/jkfn.2004.33.1.212
  3. Jung, S.L., Heung, S.S., Young, S.M., Yu, K.C., Jin, S.J. Effects of buckwheat on organ weight, glucose and lipid metabolism in streptozotocin-induced diabetic rats. J Korean Soc Food Sci Nutr 25: 831-838, 1994
  4. Choi, Y.S., Kim, B.R., Jin, L.H., Lee, B.H., Shim, T.H., Lee, S.Y. In vitro screening of dietary factors on buckwheat(Fagopyrum esculentum Moench) influencing the regulation of blood pressure, glucose and cholesterol level. J Korean Soc Food Sci Nutr 29: 280-287, 2000
  5. Ham, S.S., Choi, K.P., Choi, Y.S., Lee, S.Y. Studies on antimutagenic and lipotropic action of flavonoids of buckwheat leaf extract. J Korean Soc Food Nutr 23: 698-703, 1994
  6. Kim, Y.E., Oh, S.W., Kwon, E.K., Han, D.S., Kim, I.H., Lee, C.H. Effects of green tea, buckwheat and grape leaves extracts on lipid metabolism, antioxidative capacity, and antithrombotic activity in rats fed high cholesterol diets. Korean J Food Sci Technol 36: 979-985, 2004
  7. Tomomi, M., Buxiang, S., Aya, I. Antioxidant activies of buckwheat hull extract toward various oxidative stress in vitro and in vivo. Bio pharm Bull 24: 209-213, 2001 https://doi.org/10.1248/bpb.24.209
  8. Kim, C.D., Lee, W.K., No, K.O., Park, S.K., Lee, M.H., Lim, S.R., Roh, S.S. Anti-allergic action of buckwheat (Fagopyrum esculentum Moench) grain extract. Int Immunopharmacol 3: 129-136, 2003 https://doi.org/10.1016/S1567-5769(02)00261-8
  9. 손호용, 권정숙, 손건호, 권기석, 류희영, 금은주. 메밀 종자의 항트롬빈 활성과 혈전증 예방효과, 한국식품영양과학회지 35(2):132-138, 2006 https://doi.org/10.3746/jkfn.2006.35.2.132
  10. Ferguson, C.A. and Kidson, S.H. The regulation of tyrosinase gene transcription. Pigment Cell Res. 10: 127-138, 1997 https://doi.org/10.1111/j.1600-0749.1997.tb00474.x
  11. Hearing, V.J. and Tsukamoto, K. Enzymatic control of pigmentation in mammals. FASEB J. 5: 2902-2909, 1991 https://doi.org/10.1096/fasebj.5.14.1752358
  12. Tsukamoto, K., Jackson, I.J., Urabe, K., Montague, P.M. and Hearing, V.J. A second tyrosinase-related protein, TRP-2, is a melnogenic enzyme termed DOPAchrome tautomerase. EMBO J. 11: 519-526, 1992
  13. Boissy, R.E., Sakai, C., Zhao, H., Kobayashi, T. and Hearing, V.J. Human tyrosinase related protein-1 (TRP-1) does not function as a DHICA oxidase activity in contrast to murine TRP-1. Exp. Dermatol. 7: 198-204, 1998 https://doi.org/10.1111/j.1600-0625.1998.tb00324.x
  14. Jimbow, K., Alena, F., Dixon, W. and Hara, H. Regulatory factors of pheo- and eumelanogenesis in melanogenic compartments, pigment Cell Res. 2: 36-42, 1992
  15. Im, S., Moro, O., Peng, F., Medrano, E.E., Cornelius, J., Babcock, G., Nordlund, J.J. and Abdel-Malek, Z.A. Activation of the cAMP pathway by ${\alpha}$-melanotropin mediates the response of human melanocytes to ultraviolet B radiation. Cancer Res. 58: 47-54, 1998
  16. Kadekaro, A.L., Kanto, H., Kavanagh, R. and Abdel-Malek, Z.A. Significance of the melanocortin 1 receptor in regulating human melanocyte pigmentation, proliferation, and survival. Ann. N. Y. Acad. Sci. 994: 359-365, 2003 https://doi.org/10.1111/j.1749-6632.2003.tb03200.x
  17. Romero-Graillet, C., Aberdam, E., Biagoli, N., Massabni, W., Ortonne, J.P. and Ballotti, R. Ultraviolet B radiation acts through the nitric oxide and cGMP signal transduction pathway to stimulate melanogenesis in human melanocytes. J. Biol. Chem. 271: 28052-28056, 1996 https://doi.org/10.1074/jbc.271.45.28052
  18. Maeda, K., Fukuda, M. In vivo effectiveness of several whitening cosmetic components in human melanocytes. J. Soc. Cosmet. Chem. 42: 361-368, 1991
  19. Mosmann, T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J. Immun. Methods 65: 55-63, 1983 https://doi.org/10.1016/0022-1759(83)90303-4
  20. Matinez-Esparza, M. Mechanisms and melanogenesis inhibition by tumor necrosis factor in B16/F10 mouse melanoma cells. Eur. J. Biochem. 225: 139-146, 1998
  21. Hosoi, J., Abe, E., Suda, T., Kuroki, T. Regulation of melanin synthesis of B16 mouse melanoma cells by 1 alpha, 25-dihydroxyvitamin D3 and retinoic acid. Cancer Research 45: 1474-1478, 1985
  22. Chakraborty, A.K., Funasaka, Y., Slominski, A., Ermak, G., Hwang, J., Pawelek, J.M., Ichihasi, M. Production and release of proopiomelanocortin (POMC) derived peptides by human melanocytes and keratinocytes in culture: regulation by ultraviolet B. Biochim Biophys. Acta 1313: 130-138, 1996 https://doi.org/10.1016/0167-4889(96)00063-8
  23. Walter, E., Roger, R., Monique, D.C., Dominique, L., Jean-paul, O., Robert, B. Mitogen-activated protein kinase pathway and AP-1 are activated during cAMP-induced melanogenesis in B16 melanoma cells. J. Bio. Chem. 270: 24315-24320, 1995 https://doi.org/10.1074/jbc.270.41.24315
  24. Hearing, V.J., Tsukamoto, K., Urabe, K. et al. Functional properties of cloned melanogenic proteins. Pigment Cell Res. 5: 264-270, 1992 https://doi.org/10.1111/j.1600-0749.1992.tb00547.x
  25. Mas, J.S., Gerritsen, I., Hahmann, C., Jimenez-Cervantes, C. and Garcia-Borron, J.C. Rate-limiting factors in melanocortin 1 receptor signalling through the cAMP pathway. Pigment Cell Res. 16: 540-547, 2003 https://doi.org/10.1034/j.1600-0749.2003.00073.x
  26. Busca, R. and Ballotti, R. Cyclic AMP: a key messenger in the regulation of skin pigmentation. Pigment Cell Res. 13: 60-69, 2000 https://doi.org/10.1034/j.1600-0749.2000.130203.x
  27. Ortonne, J.P. Photoprotective properties of skin melanin. British J. Derma. 146(61):7-10, 2002
  28. Prota, G. Recent advance in the chemistry of melanogenesis in mammals. J. Invest. Dermatol. 75: 122-129, 1980
  29. Hunt, G., Donatien, P.D., Lunec, J. Cultured human melanocytes respond to MSH peptides and ACTH. Pigment Cell Res. 7: 217-221, 1994 https://doi.org/10.1111/j.1600-0749.1994.tb00052.x
  30. Im, S., Kim, J., On, W.Y., Kang, W.H. Increased expression of ${\alpha}$-Melanocyte-stimulating hormone in the lesionl skin of melasma. Br. J. Dermatol 146: 165-167, 2000
  31. Friedman, P.S., Wren, F., Buffey, J. ${\alpha}$-MSH causes a small rise in cAMP but has no effect on basal or ultraviolet-stimulated melanogenesis in human melanocytes. Br J Dermatol. 123: 145-151, 1990 https://doi.org/10.1111/j.1365-2133.1990.tb01841.x
  32. Hun, G., Todd, C., Cresswell, J.E. $\alpha$-melanocyte stimulating hormone and its analogue Nle4DPhe7 ${\alpha}$-MSH affect morphology, tyrosinase activity and melanogenesis in cultured human melanocytes. J Cell Sci. 107: 205-211, 1994
  33. Provance, D.W., Jr, Wei, M., Ipe, V., Mercer, J.A. cultured melanocytes from dilute mutant mice exhibit dendritic morphology and altered melanosome distribution. Proc Natl Acad Sci USA 93: 14554-14558, 1996
  34. Hara, M., Yaar, M., Byers, H.R., Goukassian, D., Fine, R.E., Gonsalves, J., Gilchrest, B.A. Kinesin participates in melanosomal movement along melanocyte dendrites. J Invest Dermatol. 114: 438-443, 2000 https://doi.org/10.1046/j.1523-1747.2000.00894.x
  35. Wu, X., Bowers, B., Rao, K., Wei, Q., Hammer, J.A. Visualization of melanosome dynamics within wild-type and dilute melanocytes suggests a paradigm for myosin V function In vivo. J Cell Biol. 143: 1899-1918, 1998 https://doi.org/10.1083/jcb.143.7.1899