Microbiology and Biotechnology Letters (한국미생물·생명공학회지)

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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The Effect of Genistein on Melanin Synthesis and In vivo Whitening

제니스테인의 멜라닌 생성 억제 및 In vivo 미백 효과

  • Yang, Eun-Soon (Department of Bioengineering, Graduate School at KonKuk University) ;
  • Hwang, Jae-Sung (Amorepacific Corporation, R&D Center) ;
  • Choi, Hyun-Chung (Amorepacific Corporation, R&D Center) ;
  • Hong, Ran-Hi (Department of Preventive Medicine, College of Medicine, ChungAng University) ;
  • Kang, Sang-Mo (Department of Bioengineering, Graduate School at KonKuk University)
  • 양은순 (건국대학교 대학원 생물공학과) ;
  • 황재성 (㈜아모레퍼시픽 기술연구원) ;
  • 최현정 (㈜아모레퍼시픽 기술연구원) ;
  • 홍란희 (중앙대학교 의과대학 예방의학교실) ;
  • 강상모 (건국대학교 대학원 생물공학과)
  • Published : 2008.03.28
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Abstract

The effect of genistein on melanin synthesis was studied using in vitro and in vivo model. Genistein inhibited melanin synthesis in cultured melan-a cells dose dependently. Tyrosinase activity was decreased by genistein treatment in melan-a cells, but genistein did not inhibit tyrosinase directly. Genistein did not affect the expression of tyrosinase in melan-a cells. Genistein inhibited the activity of ${\alpha}$-glucosidase in virtro and the glycosylation of tyrosinase in melan-a cells. The resulting unsaturated glycosylation of tyrosinase makes it unstable and disturb correct transportation. To further clarify the effect of genistein on the melanogenesis, we established UVB-induced hyperpigmentation on the shaved backs of brown guinea pigs. The animals were exposed to UVB radiation once a week for three consecutive weeks. Genistein (1 and 2%) or vehicle alone as a control were then topically applied to the hyperpigmented areas daily. Genistein showed significant lightening effect on the UVB-induced hyperpigmentation in five weeks. Depigmenting effect was prominent in 2% genistein treatment with Fontana-Masson staining. In conclusion, genistein may be a useful agent for skin whitening.

제니스테인의 미백제로서 응용 가능성을 알아보기 위해 멜라닌 생성에 미치는 영향을 in vitro 및 in vivo 실험을 통해 알아보았다. Melan-a 세포에 제니스테인을 처리하여 멜라닌 양을 측정한 결과 멜라닌 생성이 농도 의존적으로 감소되었다. 멜라닌 생성 억제가 tyrosinase의 활성 저해와 관련되는지를 확인해보고자 하였다. 그 결과 제니스테인은 tyrosinase에 직접 작용하여 활성을 저해하지는 않았으나, 세포내의 tyrosinase의 활성은 농도 의존적으로 저해하였다. 제니스테인이 tyrosinase의 발현에는 영향을 미치지 않았는데 이는 세포내 tyrosinase의 활성 저해가 발현과는 다른 기전에 의해 일어난다는 것을 의미한다. 그리고 제니스테인은 ${\alpha}$-glucosidase를 저해하였으며, 이를 통해 N-linked glycoprotein인 tyrosinase의 glycosylation을 저해하여 tyrosinase의 세포내 이동이나 활성을 억제할 수 있음을 알 수 있었다. 또한 제니스테인은 brown guinea pig에서 자외선에 의해 유도되는 피부흑화를 농도 의존적으로 개선하는 미백 효과가 있었다. 인공색소반에 제니스테인을 1%, 2% 도포한 결과 5주차에 대조군과 비교하여 유의적인 미백 효과를 확인할 수 있었다. 멜라닌에 대한 F-M 염색 결과를 살펴보면, 제니스테인 2%를 도포한 부위의 멜라닌 함량이 대조군 도포 부위에 비해 상당히 감소하였음을 확인할 수 있었다. 이상의 결과에서 제니스테인은 미백제로서 유용하게 활용할 가치가 있는 것으로 사료된다.

Keywords

References

  1. Afaq, F. and H. Mukhtar. 2006. Botanical antioxidants in the prevention of photocarcinogenesis and photoaging. Exp. Dermatol. 15: 678-684 https://doi.org/10.1111/j.1600-0625.2006.00466.x
  2. Alaluf, S., A. Heath, N. Carter, D. Atkins, H. Mahalingam, K. Barrett, R. Kolb, and N. Smit. 2001. Variation in melanin content and composition in type V and VI photoexposed and photoprotected human skin: the dominant role of DHI. Pigment Cell Res. 14: 337-347 https://doi.org/10.1034/j.1600-0749.2001.140505.x
  3. Ando, H., A. Ryu, A. Hashimoto, M. Oka, and M. Ichashi. 1998. Linoleic and $\alpha$-linoleic acid lightens ultravioletinduced hyperpigmentation of the skin. Arch. Dermatol. Res. 290: 375-381 https://doi.org/10.1007/s004030050320
  4. Angelov, A., M. Putyrski, and W. Liebl. 2006. Molecular and biochemical characterization of alpha-glucosidase and alpha-mannosidase and their clustered genes from the thermoacidophilic archaeon Picrophilus torridus. J. Bacteriol. 188: 7123-7131 https://doi.org/10.1128/JB.00757-06
  5. Battaini, G., E. Monzani, L. Casella, L. Santagostini, and R. Pagliarin. 2000. Inhibition of the catecholase activity of biomimetic dinuclear copper complexes by kojic acid. J. Biol. Inorg. Chem. 5: 262-268 https://doi.org/10.1007/s007750050370
  6. Bennett, D. C., P. J. Cooper, and I. R. Hart. 1987. A line of non-tumorigenic mouse melanocytes, syngeneic with the B16 melanoma and requiring a tumour promoter for growth. Int. J. Cancer 39: 414-418 https://doi.org/10.1002/ijc.2910390324
  7. Bolognia, J. L., M. S. Murray, and J. M. Pawelek. 1990. Hairless pigmented guinea pigs: a new model for the study of mammalian pigmentation. Pigment Cell Res. 3: 150-156 https://doi.org/10.1111/j.1600-0749.1990.tb00280.x
  8. Branza-Nichita, N., A. J. Petrescu, R. A. Dwek, M. R. Wormald, F. M. Platt, and S. M. Petrescu. 1999. Tyrosinase folding and copper loading in vivo: a crucial role for calnexin and alpha-glucosidase II. Biochem. Biophys. Res. Commun. 261: 720-725 https://doi.org/10.1006/bbrc.1999.1030
  9. Branza-Nichita, N., G. Negroiu, A. J. Petrescu, E. F. Garman, F. M. Platt, M. R. Wormald, R. A. Dwek, and S. M. Petrescu. 2000. Mutations at critical N-glycosylation sites reduce tyrosinase activity by altering folding and quality control. J. Biol. Chem. 275: 8169-8175 https://doi.org/10.1074/jbc.275.11.8169
  10. Briganti, S., E. Camera, and M. Picardo. 2003. Chemical and Instrumental Approaches to Treat Hyperpigmentation. Pigment Cell Res. 16: 101-110 https://doi.org/10.1034/j.1600-0749.2003.00029.x
  11. Chang, T. S., H. Y. Ding, and H. C. Lin. 2005. Identifying 6,7,4'-trihydroxyisoflavone as a potent tyrosinase inhibitor. Biosci. Biotechnol. Biochem. 69: 1999-2001 https://doi.org/10.1271/bbb.69.1999
  12. Choi, H., J. Lee, H. J. Shin, B. G. Lee, I. Chaing, and J. S. Hwang. 2004. Deoxypodophyllotoxinreduces skin pigmentation of brown guinea pigs. Planta. Med. 70: 378-380 https://doi.org/10.1055/s-2004-818955
  13. Choi, H., S. Ahn, H. Chang, N. S. Cho, K. Joo, B. G. Lee, I. Chang, and J. S. Hwang. 2007. Influence of N-glycan processing disruption on tyrosinase and melanin synthesis in HM3KO melanoma cells. Exp. Dermatol. 16: 110-117 https://doi.org/10.1111/j.1600-0625.2006.00515.x
  14. Franchi, J. 2000. Depigmenting effects of calcium D-pantetheine- S-sulfonate on human melanocytes. Pigment Cell Res. 13: 165-171 https://doi.org/10.1034/j.1600-0749.2000.130308.x
  15. Fuller, B. B., J. B. Lunsford, and D. S. Iman. 1987. Alphamelanocyte- stimulating hormone regulation of tyrosinase in Cloudman S-91 mouse melanoma cell cultures. J. Biol. Chem. 262: 4024-4033
  16. Goochee, C. F. 1992. Bioprocess factors affecting glycoprotein oligosaccharide structure. Dev. Biol. Stand. 76: 95-104
  17. Halaban, R. E., E. Cheng, Y. Zhang, G. Moellmann, D. Hanlon, M. Michalak, V. Setaluri, and D. N. Hebert. 1997. Aberrant retention of tyrosinase in the endoplasmic reticulum mediates accelerated degradation of the enzyme and contributes to the dedifferentiated phenotype of amelanotic melanoma cells. Proc. Natl. Acad. Sci. USA. 94: 6210-6215
  18. Hwang, J. S., H. Shin, H. S. Noh, H. Choi, S. Ahn, D. S. Park, D. H. Kim, B. G. Lee, I. Chang, and H. H. Kang. 2002. The inhibitory effects of 3,4,5-Trimethoxy cinnamate thymol ester(TCTE, Melasolv) on Melanogenesis. Journal of the Society of Cosmetic Scientists of Korea. 28: 135-149
  19. Imokawa, G. and Y. Mishima. 1984. Functional analysis of tyrosinase isozymes of cultured malignant melanoma cells during the recovery period following interrupted melanogenesis induced by glycosylation inhibitors. J. Invest. Dermatol. 83: 196-201 https://doi.org/10.1111/1523-1747.ep12263562
  20. Kim, D. S., S. Y. Kim, S. J. Moon, J. H. Chung, K. H. Kim, K. H. Cho, and K. C. Park. 2001. Ceramide inhibits cell proliferation trough Akt/PKB inactivation and decreases melanin synthesis in Mel-Ab Cells. Pigment Cell Res. 14: 110-115 https://doi.org/10.1034/j.1600-0749.2001.140206.x
  21. Kornfeld, R. and S. Kornfeld. 1985. Assembly of asparaginelinked oligosaccharides. Annu. Rev. Biochem. 54: 631-664 https://doi.org/10.1146/annurev.bi.54.070185.003215
  22. Lee, D. S. and S. H. Lee. 2001. Genistein, a soy isoflavone, is a potent alpha-glucosidase inhibitor. FEBS Lett. 501: 84-86 https://doi.org/10.1016/S0014-5793(01)02631-X
  23. Lee, K. H. 2004. Antioxidantive properties of isoflavone, genistein from soybean and its application to whitening cosmetics, M.S. Thesis, Seoul National University of Technology, Seoul
  24. Maeda, K. and M. Fukuda. 1996. Arbutin: mechanism of its depigmenting action in human melanocyte culture. J. Pharmacol. Exp. Ther. 276: 765-769
  25. Mishima, Y. and G. Imokawa. 1983. Selective aberration and pigment loss in melanosomes of malignant melanoma cells in vitro by glycosilation inhibitors: premelanosomes as glycoprotein. J. Invest. Dermatol. 81: 106-114 https://doi.org/10.1111/1523-1747.ep12542192
  26. Petrescu, A. J., T. D. Butters, G. Reinkensmeier, S. Petrescu, F. M. Platt, R. A. Dwek, and M. R. Wormal. 1997. The solution NMR structure of glucosylated N-glycans involved in the early stages of glycoprotein biosynthesis and folding. EMBO J. 16: 4302-4310 https://doi.org/10.1093/emboj/16.14.4302
  27. Petrescu, S. M., A. J. Petrescu, H. N. Titu, R. A. Dwek, and F. M. Platt. 1997. Inhibition of N-glycan processing in B16 melanoma cells results in inactivation of tyrosinase but does not prevent its transport to the melanosome. J. Biol. Chem. 272: 15796-15803 https://doi.org/10.1074/jbc.272.25.15796
  28. Rauth, S., J. Kichina, and A. 1997. Green. Inhibition of growth and induction of differentiation of metastatic melanoma cells in vitro by genistein: chemosensitivity is regulated by cellular p53. Br. J. Cancer 75: 1559-1566 https://doi.org/10.1038/bjc.1997.268
  29. Wang, Y., L. Ma, C. Pang, M. Huang, Z. Huang, and L. Gu. 2004. Synergetic inhibition of genistein and D-glucose on $\alpha$- glucosidase. Bioorg. Med. Chem. Lett. 14: 2947-2950 https://doi.org/10.1016/j.bmcl.2004.03.035
  30. Wang, Y., L. Ma, Z. Li, Z. Du, Z. Liu, J. Qin, X. Wang, Z. Huang, L. Gu, and A. S. Chen. 2004. Synergetic inhibition of metal ions and genistein on alpha-glucosidase. FEBS Lett. 576: 46-50 https://doi.org/10.1016/j.febslet.2004.08.059
  31. Wei, H., R. Bowen, X. Zhang, and M. Lebwohl. 1998. Isoflavone genistein inhibits the initiation and promotion of two-stage skin carcinogenesis in mice. Carcinogenesis. 19: 1509-1514 https://doi.org/10.1093/carcin/19.8.1509
  32. Wei, H., X. Zhang, Y. Wang, and M. Lebwohl. 1993. Inhibition of tumor promoter-induced hydrogen peroxide formation in vitro and in vivo by genistein. Nutr. Cancer. 20: 1-12 https://doi.org/10.1080/01635589309514265
  33. Wei, H., X. Zhang, Y. Wang, and M. Lebwohl. 2002. Inhibition of ultraviolet light-induced oxidative events in the skin and internal organs of hairless mice by isoflavone genistein. Cancer Lett. 185: 21-29 https://doi.org/10.1016/S0304-3835(02)00240-9
  34. Winchester, B. and G. W. Fleet. 1992. Amino-sugar glycosidase inhibitors: versatile tools for glycobiologists. Glycobiology 2: 199-210 https://doi.org/10.1093/glycob/2.3.199
  35. Yan, C. H., X. G. Chen, Y. Li, and R. Han. 1999. Effects of genistein, a soybean-derived isoflavone, on proliferation and differentiation of B16-BL6 mouse melanoma cells. J. Asian Nat. Prod. Res. 1: 285-299 https://doi.org/10.1080/10286029908039877