DOI QR코드

DOI QR Code

Anti-oxidant Function and Inhibitory Effects of the Expression of MITF, TRP-1, TRP-2 and Tyrosinase of Sesamum indicum L. in B16F10 Melanoma Cells

참깨 추출물의 항산화 활성 및 melanoma cell (B16F10)에서 MITF, TRP-1, TRP-2, tyrosinase 의 발현 저해

  • Yoo, Dan-Hee (Department of Herbal Cosmetic Science, Hoseo University) ;
  • Joo, Da-Hye (Department of Herbal Cosmetic Science, Hoseo University) ;
  • Lee, Jin-Young (Department of Herbal Cosmetic Science, Hoseo University)
  • 유단희 (호서대학교 한방화장품과학과) ;
  • 주다혜 (호서대학교 한방화장품과학과) ;
  • 이진영 (호서대학교 한방화장품과학과)
  • Received : 2016.10.13
  • Accepted : 2017.02.13
  • Published : 2017.03.30

Abstract

This study was performed to improve the antioxidant and skin-whitening activities of 70% ethanol extract from Sesamum indicum L. (SIL). The electron-donating ability of the SIL extract was 71.7% at a concentration of $1,000{\mu}g/ml$. The whitening effects that was measured by tyrosinase inhibition assay. As a result, SIL extract was shown 42% at $1,000{\mu}g/ml$ concentration. The cell toxicity on B16F10 melanoma cells of SIL of 70% ethanol extract showed 84.3% at $1,000{\mu}g/ml$ concentration. The microphthalmia-associated transcription factor (MITF), tyrosinase relate protein-1 (TRP-1), tyrosinase relate protein-2 (TRP-2) and Tyrosinase protein and mRNA expression inhibitory effect of SIL extract were measured by western blot and reverse transcription- polymerase chain reaction (PCR) at 50, 250, $500{\mu}g/ml$ concentration. Consequently, the MITF, TRP-1, TRP-2, Tyrosinase protein expression inhibitory effect of SIL extract was decreased by 68.3%, 39.2%, 89.7%, 22.3%, respectively, at $500{\mu}g/ml$ concentration. Moreover, MITF, TRP-1, TRP-2, Tyrosinase mRNA expression inhibitory effect by reverse-transcription-PCR of SIL extract was decreased by 81.8%, 66.5%, 84.2%, 68.1%, respectively, at $500{\mu}g/ml$ concentration. Therefore, we excellently identified the antioxidant activities and whitening effect of SIL extract, and this finding suggested that SIL extract has great potential as a cosmetic ingredients.

본 연구는 참깨 70% 에탄올 추출물의 항산화 효과와 미백효과를 검증하여 나타내었다. 참깨 추출물의 전자공여능 측정실험은 $1,000{\mu}g/ml$에서 71.7%의 효과를 나타냈으며, tyrosinase 저해활성 측정은 $1,000{\mu}g/ml$의 농도에서 42%의 효과를 보였다. 참깨 추출물의 세포 생존율을 melanoma cell (B16F10)에서 확인하기 위하여 MTT assay를 진행하였으며, 세포 생존율을 측정한 결과, $1,000{\mu}g/ml$ 농도에서 84.3%의 독성을 보였다. 이하의 세포실험에서는 세포 생존율이 90% 이상되는 농도인 $500{\mu}g/ml$ 이하에서 실험을 진행하였다. 참깨 추출물의 MITF, TRP-1, TRP-2 및 tyrosinase의 단백질 발현 효과를 50, 250, $500{\mu}g/ml$ 농도에서 측정하였으며, 그 결과 MITF, TRP-1, TRP-2 및 tyrosinase의 각각 $500{\mu}g/ml$ 농도에서 68.3%, 39.2%, 89.7%, 22.3%의 단백질 발현 억제 효과를 보였다. 또한, MITF, TRP-1, TRP-2 및 tyrosinase의 mRNA발현을 RT-PCR로 50, 250, $500{\mu}g/ml$ 농도에서 측정하였고, 양성 대조군으로 GAPDH를 사용하였다. 그 결과, 참깨 추출물의 $500{\mu}g/ml$ 농도에서 각각 81.8%, 66.5%, 84.2%, 68.1%의 mRNA 발현이 감소함을 확인할 수 있었다. 이상의 결과로 부터 참깨 추출물의 화장품 소재로서의 가능성을 확인할 수 있었다.

Keywords

References

  1. Ashri, A. 1998. Sesame breeding. Plant Breed. Rev. 16, 179-228.
  2. Blois, M. S. 1958. Antioxidant determination by the use of a stable free radical. Nature 26, 1199-1120.
  3. Carmichael, J., DeGraff, W. G., Gazdar, A. F., Minna, J. D. and Mitchell, H. B. 1987. Evaluation of a tetrazolium based semiautomated colorimetric assay: assessment of chemosensitivity testing. Cancer Res. 47, 936-942.
  4. Cho, J. H., Kim, H. J. and Kim, Y. B. 2003. The study on depigmentation effects of salt, acetum, sesami semen nigrum. J. Kor. Med. Ophthalmol. Otolaryngol. Dermatol. 16, 79-95.
  5. Han, G. S., Kim, D. S., Woo, W. H. and Mun, Y. J. 2010. Inhibitory effect of Fructus ligustri lucidi on tyrosinase and MITF expressions. Kor. J. Ori. Physiol. Pathol. 24, 296-301.
  6. Hearing, V. J. 1999. Biochemical control of melanogenesis and melanosomal organization. J. Investig. Dermatol. Symp. Proc. 4, 24-28. https://doi.org/10.1038/sj.jidsp.5640176
  7. Hwang, J. and Lee, B. M. 2007. Inhibitory effects of plant extracts on tyrosinase, L-DOPA oxidation, and melanin synthesis. J. Toxicol. Environ. Health A. 70, 393-407. https://doi.org/10.1080/10937400600882871
  8. Jin, K. S., Oh, Y. N., Park, J. A., Lee, J. Y., Jin, S. J., Hyun, S. K., Hwang, H. J., Kwon, H. J. and Kim, B. W. 2012. Anti-oxidant, anti-melanogenic, and anti-inflammatory activities of Zanthoxylum schinifolium extract and its solvent fractions. Ko. J. Microbiol. Biotecnol. 40, 317-379.
  9. Kamal Eldin, A. and Appelqvist, L. A. 1994. Variation in fatty acid composition of the different acyl lipids in seed oils from four Sesamum species. J. Am. Chem. Soc. 71, 135-139.
  10. Kikugawa, K., Arai, M. and Kurechi, T. 1983. Participation of sesamol in stability of sesame oil. J. Am. Chem. Soc. 60, 1528-1533.
  11. Kim, D. H., Moon, Y. S. and Son, J. H. 2015. Verification of anti-oxidative activity of Aruncus dioicus, a native plant of Ulleungdo. J. Plant Biol. 42, 55-59.
  12. Kim, H. H., Park, G. H., Park, K. S., Lee, J. Y., Kim, T. H. and An, B. J. 2010. The effects of Aster glehni Fr. Schm. Extracts on whitening and anti-wringkle. J. Life Sci. 20, 1034-1040. https://doi.org/10.5352/JLS.2010.20.7.1034
  13. Lin, C. B., Babiarz, L., Liebel, F., Roydon-Price, E., Kizoulis, M., Gendimenico, G. J., Fisher, D. E. and Seiberg, M. 2002. Modulation of microphthalmia-associated transcription factor gene expression alters skin pigmentation. J. Invest. Dermatol. 119, 1330-1340. https://doi.org/10.1046/j.1523-1747.2002.19615.x
  14. Makinde, F. M. and Akinoso, R. 2014. Comparison between the nutritional quality of flour obtained from raw, roasted and fermented sesame (Sesamum indicum L.) seed grown in Nigeria. Acta. Sci. Pol. Technol. Aliment. 13, 309-319. https://doi.org/10.17306/J.AFS.2014.3.9
  15. Mallick, S., Singh, S. K., Sarkar, C., Saha, B. and Bhadra, R. 2005. Human placental lipid induces melanogenesis by increasing the expression of tyrosinase and its related proteins in vitro. Pigment Cell Res. 18, 25-33. https://doi.org/10.1111/j.1600-0749.2004.00193.x
  16. Mullany, L. C., Darmstradt, G. L., Khatry, S. K. and Titlsch, J. M. 2005. Traditional massage of newborns in Nepal: implications for trials of improved practice. J. Trop. Pediatr. 51, 82-86. https://doi.org/10.1093/tropej/fmh083
  17. Park, C. G. 2002. Korean Oriental Medicine special lecture, Jipmoondang, Korea.
  18. Park, S. H., Kim, A. H., Park, B. J. and Kim, J. J. 2013. Whitening effect of Biochanin A. J. Soc. Cosmet. Sci. Korea 39, 289-294.
  19. Seo, E. J., Hong, E. S., Choi, M. H., Kim, K. S. and Lee, S. J. 2010. Antioxidant and skin whitening effects of Ramnus yoshinoi extracts. Kor. J. Food Sci. Technol. 42, 750-754.
  20. Son, H. U., Lee, S. H., Kim, M. A., Parl, H. J. and Lee, S. H. 2012. Comparison of melanogenesis-inhibiting activity by extracts of Prunus persica flower and Calyx. Kor. J. Food Preserv. 19, 946-950. https://doi.org/10.11002/kjfp.2012.19.6.946
  21. Ukeda, H., Maeda, S., Ishii, T. and Sawamura, M. 1997. Spectrophotometric assay for superoxide dismutase based on tetrazolium salt 3'-1-(phenylamino)-carbonyl-3,4-tetrazolium]-bis(4-methoxy-6-nitro) benzenesulfonic acid hydrate reduction by xanthine-xanthine oxidase. Anal. Biochem. 251, 206- 209. https://doi.org/10.1006/abio.1997.2273
  22. Yagi, A., Kanbara, T. and Morinobu, N. 1987. Inhibition of mushroom-tyrosinase by aloe extract. Planta Med. 53, 515-517. https://doi.org/10.1055/s-2006-962798