Journal of the Korea Academia-Industrial cooperation Society (한국산학기술학회논문지)

The Korea Academia-Industrial cooperation Society (한국산학기술학회)
권호 표지
DOI QR코드

DOI QR Code

Whitening Effects of Angelica dahurica Radix Ethanol Extract

백지 에탄올추출물의 미백효능 연구

  • Kim, Pil-Sun (Dept. of Herbal Skin Care, Daegu Haany University)
  • 김필순 (대구한의대 한방피부미용학과)
  • Received : 2011.07.21
  • Accepted : 2011.09.08
  • Published : 2011.09.30
Download PDF
⟨ Previous Next ⟩

Abstract

To investigate whitening Effects of Angelica dahurica Radix Ethanol Extract (ADEE), we used melan-a cell line, brown guinea pig, and HMB-45. We treated with ADEE of 6.25, 12.5, 25, and 50 ${\mu}g/m{\ell}$ concentration in order to evaluate the effect of ADEE on cell viability and on morphological observation of melan-a cells. Also we were induced the artificial tanning spots by 1,500 mJ/$cm^2$ of ultraviolet B radiation on the backs of brown guinea pigs (approximately 450~500g) and then the test agent of $30{\mu}{\ell}$ was applied on the spots twice a day, five days a week, for five weeks respectively. The visible whitening effect was evaluated once a week. At the end of the experiment, the animals were sacrificed under anesthetization. The artificial tanning spots were obtained by biopsy punch and stained with HMB-45 to observe the gp100 proteins which were melanosomes. Our results show that cell viability was not reduce at ADEE concentrations between 6.25 and 50 ${\mu}g/m{\ell}$, melanin synthesis and melanocyte dendricity were decreased in ADEE treated melan-a cells increasing ADEE concentration. In the gross observation, ADEE treated groups had lower pigmentation than the vehicle control groups. And in the histological observation, ADEE treated groups had lower melanocytes than the vehicle control groups. Also in the quantitative analysis of the gp100 proteins using image analysis software, ADEE treated groups had a significantly lower value (p<0.001) than the vehicle control group and this resultsagreed with the results of observation under microscope. From these results, weconcluded that ADEE had positive whitening effect.

본 논문은 백지 에탄올추출물의 미백효능을 조사하기 위하여 melan-a cell, 브라운 기니피그 및 HMB-45 염색을 사용하였다. melan-a 세포에 시료를 6.25, 12.5, 25, 50, ${\mu}g/m{\ell}$ 농도로 처치하여 세포 생장율의 변화 관찰 및 멜라닌세포의 형태학적 변화를 관찰하였다. 또한 brown guinea pig (450~500 g)등 부위에 1500 mJ/$cm^2$ 광량의 ultraviolet B조사로 유발시킨 인공색소반 (${\Phi}$ 2 mm)에 1일 2회, 주 5일, 매회 30 ${\mu}{\ell}$씩 총 5주간 시료를 도포하여 주 1회 육안적 변화를 관찰 하였고, 실험 5주째 되는 날 실험동물은 염산 케타민으로 마취 후 시료를 도포한 인공색소반 부위를 biopsy punch로 절취하여 HMB-45 염색으로 멜라닌소체 내 glycoprotein(gp100) 단백질의 조직학적 변화를 관찰 하였다. 세포생장율 측정에서 시료처치 6.25~50 ${\mu}g/m{\ell}$에서의 세포생장율은 높았고, 멜라닌세포의 형태학적 변화에서는 시료처치 농도가 증가할수록 멜라닌합성 및 수지상 돌기가 적게 관찰 되었다. 동물 실험 육안적 관찰에서 시료 도포군이 용매대조군에 비해 멜라닌 색소침착 정도가 확연하게 밝아졌고, gp100 단백질의 조직학적 관찰에서 시료 도포군은 gp100 단백질 발현 정도가 현저하게 줄어들었으며, 영상분석에서도 시료 도포군이 용매대조군에 비해 유의하게(p<0.001) 감소하였는데 이러한 결과는 현미경 관찰 결과와 일치하였다. 이상의 결과를 종합하면 백지 에탄올추출물은 우수한 미백효과가 있는 것으로 판단된다.

Keywords

References

  1. Y. Matsumura, and H. N. Ananthaswamy, "Toxic effects of ultraviole radiation on the skin", Toxicology and Applied Pharmacology, Vol. 195, pp. 298-308, 2004. https://doi.org/10.1016/j.taap.2003.08.019
  2. G. P. He, et al., "In vivo measurement and mapping of skin redox stress induced by ultra violet light exposure", Free Radical Biology & Medicine, Vol. 36, no. 5, pp. 665-672, 2004. https://doi.org/10.1016/j.freeradbiomed.2003.11.024
  3. S. H. Jung, et al., "Inhibitory Effects of fucoidan on Melanin Synthesis and Tyrosinase Activity", Journal of Life Science, Vol. 19, no. 1, pp. 75-80, 2009. https://doi.org/10.5352/JLS.2009.19.1.075
  4. K. H. Jang, and S. S. Roh, "Depigmentation activity of many herb", Research Institute of Korean Medicine, vol. 13, no. 2, pp. 289-302. 2004.
  5. E. K. Sung, et al, "The investigation of the melanocytes in a cultured skin equivalent model",Korean Journal of Anatomy, Vol. 33, no. 1, pp. 89-97, 2000.
  6. B. B. Fuller, et al., "Regulation of catalytic activity of prxisting tyrosinase in Black and Caucasian human melanocyte cell cultures", Experimental Cell Research, Vol. 262, pp.197-208, 2001. https://doi.org/10.1006/excr.2000.5092
  7. N. Agar, and A. R. Young, "Melanogenesis: a photoprotective response to DNA damage?", Mutation Research, Vol. 571, no. 1, pp. 121-132, 2005. https://doi.org/10.1016/j.mrfmmm.2004.11.016
  8. K. Wakamatsu, et al., "Usefulness of 4-amino-3-hydroxy-phenylalanine as a specific marker of pheomelanin", Pigment Cell Research, Vol. 15, pp. 225-23, 2002. https://doi.org/10.1034/j.1600-0749.2002.02009.x
  9. A. Slominski, et al., "Melanin pigmentation in mammalian skin and its hormonal regulation". Physiological Reviews, Vol. 84, no. 4, pp. 1155-1228, 2004. https://doi.org/10.1152/physrev.00044.2003
  10. S. Briganti, et al., "Chemical and instrumental approaches to treat hyperpigmentation", Pigment Cell Research, Vol. 16, no. 2, pp. 101-11, 2003. https://doi.org/10.1034/j.1600-0749.2003.00029.x
  11. K. Maeda, and M. Hatato, "Involvement of photooxidation of melanogenic precursors in prolonged pigmentation induced by ultraviolet A", Journal of Investigative Dermatology, Vol. 122, pp. 503-509, 2004. https://doi.org/10.1046/j.0022-202X.2004.22223.x
  12. J. Cabanes, et al., Kojic acid, a cosmetic skin whitening agent, is a show binding inhibitor of cathecilase activity of tyrosinase", Journal of Pharmacology, Vol. 46, no. 12, pp. 982-985, 1994. https://doi.org/10.1111/j.2042-7158.1994.tb03253.x
  13. T. Aburjai, and F. M. Natsheh, Plants used in cosmetics", Phytotherapy Research, Vol. 17, no. 9, pp. 987-1000, 2003. https://doi.org/10.1002/ptr.1363
  14. H. Y. Lee, "Study for effect of natural product extracts on skin malanogenesis inhibition", Ph.D. diss., Kangwon National University, 2009.
  15. E. Y. Joo, and W. J. Kang, "Analysis on the components of the Angelica dahurica root", Korean Journal of Food Preservation, Vol. 12, no. 5, pp. 476-481. 2005.
  16. H. S. Kim, and H. J. Chi, "Studies on essential oils of plants of Angelica genus in Korea (III)", Korean Journal of Pharmacognosy, Vol. 21, no. 2, pp. 121-125, 1990.
  17. C. M. Kim, et al., "Antithrombotic effect of the BuOH soluble fraction of Angellica dahurica root", Korean Journal of Pharmacognosy, Vol. 26, no. 1, pp. 74-77, 1995.
  18. D. Lechne, et al., "The anti-staphylococcal activity of Angelica dahurica", Phytochemistry, Vol. 65, no. 3, pp. 331-335, 2004. https://doi.org/10.1016/j.phytochem.2003.11.010
  19. M. H. Jin, et al., "Promoting synthesis ofcollagen from root", Korean Journal of Pharmacognosy, Vol. 35, no. 4, pp. 315-319, 2004.
  20. Y. S. Lee, et al., "Antioxidative activity and physiological function of the roots", Journal of Korean Society of Food Science and Nutrition, Vol. 36, no. 1, pp. 20-26, 2007. https://doi.org/10.3746/jkfn.2007.36.1.020
  21. J. L. Bolognia, et al., "Hairless pigmented guinea pigs: a new model for the study of mammalian pigmentation", Pigment Cell Research, Vol. 3, pp. 150-156, 1990. https://doi.org/10.1111/j.1600-0749.1990.tb00280.x
  22. H. J. Choi, et al. "Deoxypodophyllotoxin reduces skin pigmentation of brown guineapigs", Planta Medic, Vol. 70, pp. 378-380, 2004. https://doi.org/10.1055/s-2004-818955
  23. M. Tobiishi, et al., "Pigmentation in intrinsically aged skin of A1 guinea pigs", Pigment Cell Research, Vol. 17, no. 6, pp. 651-65, .2004. https://doi.org/10.1111/j.1600-0749.2004.00183.x
  24. J. T. Ko, et al., "Effect of Scirpi rhizoma Ethanol Extract on Skin Whitening in an Animal Model of Brown Guinea Pigs", Journal of Korean Society of Environmental Health and Toxicology, Vol. 24, no. 3, pp. 219-229, 2009.
  25. D. W. Provance, et al., "Cultured melanocytes from dilute mutant mice exhibit dendritic morphology and altered melanosome distribution", Proceedings of National Academy of Sciences USA, Vol. 93, no. 25, pp. 14554-14558, 1996. https://doi.org/10.1073/pnas.93.25.14554
  26. K. H. Son, "A study on the evaluation of depigmenting efficacy in the skin and the search for depigmenting agents from natural products", Ph.D. diss., Kangwon National University, 2007.
  27. K. Nakazawa, et al.,"Modulation of normal human melanocyte dendricity by growth promoting agents", Pigment Cell Research, Vol. 6, no. 6, pp. 406-416, 1993. https://doi.org/10.1111/j.1600-0749.1993.tb00623.x
  28. B. A. Gilchrest, et al., "Mechanisms of ultraviolet light-induced pigmentation", Journal of Photochemistry and Photobiology, Vol. 63, no. 1, pp. 1-10, 1996. https://doi.org/10.1111/j.1751-1097.1996.tb02988.x
  29. M. Seiji, et al., "Subcellular localization of melanin biosynthersi", The New York Academy of Sciences, Vol. 100, pp. 497-533, 1963. https://doi.org/10.1111/j.1749-6632.1963.tb42869.x
  30. M. V. Sheffield, et al., "Comparison of five antibodies as markers in the diagnosis of melanoma in cytologic preparations", American Journal of Clinical Pathology, Vol. 118, no. 6, pp. 930-936, 2002. https://doi.org/10.1309/EWK9-LUPR-6BC5-1GXV
  31. B. R. Smoller, et al., "HMB-45 recognizes stimulated melanocytes", Journal of Cutaneous Pathology, Vol. 16, pp. 49-53, 1989. https://doi.org/10.1111/j.1600-0560.1989.tb00010.x
  32. P. S. Kim, and Y. C. Kim, "The Whitening Effect of Angelicae dahuricae Radix Water Extract in Brow Guinea Pigs", Korean Journal of Environmental Health Science, Vol. 35, no. 5. pp. 417-425, 2009. https://doi.org/10.5668/JEHS.2009.35.5.417