Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Whitening Activity of Sambucus Sieboldiana Var. Pendula (Nakai) Extract

말오줌나무 추출물의 미백활성 검증

  • Yoo, Dan-Hee (Department of Cosmetic and Biotechnology, Hoseo University) ;
  • Kim, Jin-Tae (Department of Cosmetic and Biotechnology, Hoseo University) ;
  • Oh, Min-Jeong (Department of Cosmetic and Biotechnology, Hoseo University) ;
  • Yeom, Hyeon-Ji (Department of Cosmetic and Biotechnology, Hoseo University) ;
  • Lee, Jin-Young (Department of Cosmetic and Biotechnology, Hoseo University)
  • 유단희 (호서대학교 화장품생명공학부) ;
  • 김진태 (호서대학교 화장품생명공학부) ;
  • 오민정 (호서대학교 화장품생명공학부) ;
  • 염현지 (호서대학교 화장품생명공학부) ;
  • 이진영 (호서대학교 화장품생명공학부)
  • Received : 2018.10.23
  • Accepted : 2018.12.20
  • Published : 2019.03.30
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Abstract

This study evaluated the anti-oxidant and whitening effects of a 70% ethanol extract of the Sambucus sieboldiana var. pendula (Nakai) (SS). At $1,000{\mu}g/ml$ concentration, the electron donating ability of this SS extract was found to be 86.21% and the ABTS+ radical scavenging ability was 97.9%. In terms of whitening activity, the tyrosinase inhibitory effect of the extract was 37%, also at $1,000{\mu}g/ml$ concentration. To explore the extractefftoxicity to B16F10 melanoma cells, a 3-[4,5-dimethyl-thiazol-2-yl]-2,5-diphenyl-tetrazoliumbromide assay was performed. Results showed 90% or more cells remained viable at $100{\mu}g/ml$ concentration. A Western blot of the SS extract was used to measure microphthalmia-associated transcription factor (MITF), tyrosinase-related protein-1 (TRP-1), tyrosinase relate protein-2 (TRP-2), and the tyrosinase protein expression inhibitory effect at 25, 50, $100{\mu}g/ml$ concentrations; ${\beta}-actin$ was used as a positive control. Consequently, the MITF, TRP-1, TRP-2, and the tyrosinase protein expression inhibitory effect were seen to decrease by 34.5%, 45.6%, 58.4%, and 79.6%, respectively, at $100{\mu}g/ml$ concentration. These were also then measured by reverse transcription-polymerase chain reaction at 25, 50, $100{\mu}g/ml$ concentrations with GAPDH as a positive control. As a result, the SS extract was seen to decrease MITF, TRP-1, TRP-2, and the tyrosinase protein expression inhibitory effect by 85.4%, 67.5%, 85.2%, 67.1%, respectively at the $100{\mu}g/ml$ concentration. We therefore confirmed the possibility of Sambucus sieboldiana var. pendula (Nakai) extract as a whitening material.

본 연구는 말오줌나무 70% 에탄올 추출물의 항산화 효과와 미백 효과를 검증하였다. 말오줌나무 추출물의 전자공여능 측정실험은 $1,000{\mu}g/ml$에서 86.21%의 효과를 나타내었고, ABTS 소거능을 측정한 결과 $1,000{\mu}g/ml$ 농도에서 97.9%의 소거능을 보였다. Tyrosinase 저해활성 측정은 $1,000{\mu}g/ml$ 농도에서 37%의 효과를 보였다. 말오줌나무 추출물의 세포 생존율을 melanoma cell (B16F10)에서 확인하기 위해 MTT assay를 진행하였는데 세포 생존율을 측정한 결과, $100{\mu}g/ml$ 농도에서 90% 이상의 생존율을 보였다. 말오줌나무 추출물의 MITF, TRP-1, TRP-2 및 tyrosinase의 단백질 발현 효과를 25, 50, $100{\mu}g/ml$ 농도에서 측정하였으며 양성 대조군으로 ${\beta}-actin$을 사용하였다. 그 결과, MITF, TRP-1, TRP-2 및 tyrosinase의 각각 $100{\mu}g/ml$ 농도에서 34.5%, 45.6%, 58.4%, 79.6%의 단백질 발현 억제 효과를 보였다. MITF, TRP-1, TRP-2 및 tyrosinase의 mRNA발현을 RT-PCR로 25, 50, $100{\mu}g/ml$ 농도에서 측정하였고, 양성 대조군으로 GAPDH를 사용하였다. 그 결과, 말오줌나무 추출물의 $100{\mu}g/ml$ 농도에서 각각 85.4%, 67.5%, 85.2%, 67.1%의 mRNA 발현이 감소함을 확인 할 수 있었다. 따라서, 말오줌나무 추출물이 미백 소재로서의 가능성을 확인할 수 있었다.

Keywords

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Fig. 1. Electron donating ability of extract from Sambucus sieboldiana var. pendula (Nakai).

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Fig. 2. ABTS+ radical scavenging activity of Sambucus sieboldiana var. pendula (Nakai) extracts.

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Fig. 3. Inhibition rate of Sambucus sieboldiana var. pendula (Nakai) extracts on tyrosinase.

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Fig. 4. Cell viability of extracts from Sambucus sieboldiana var. pendula (Nakai) on melanoma cell (B16F10).

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Fig. 5. MITF protein expression rate of Sambucus sieboldiana var. pendula (Nakai) extracts on melanoma cell (B16F10).

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Fig. 6. TRP-1 protein expression rate of Sambucus sieboldiana var. pendula (Nakai) extracts on melanoma cell (B16F10).

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Fig. 7. TRP-2 protein expression rate of Sambucus sieboldiana var. pendula (Nakai) extracts on melanoma cell (B16F10).

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Fig. 8. Tyrosinase protein expression rate of Sambucus sieboldiana var. pendula (Nakai) extracts on melanoma cell (B16F10).

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Fig. 9. MITF mRNA expression rate of Sambucus sieboldiana var. pendula (Nakai) extracts on melanoma cells (B16F10).

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Fig. 10. TRP-1 mRNA expression rate of Sambucus sieboldiana var. pendula (Nakai) extracts on melanoma cells (B16F10).

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Fig. 11. TRP-2 mRNA expression rate of Sambucus sieboldiana var. pendula (Nakai) extracts on melanoma cells (B16F10).

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Fig. 12. Tyrosinase mRNA expression rate of Sambucus sieboldiana var. pendula (Nakai) extracts on melanoma cells (B16F10).

Table 1. Sequence of the primers used for RT-PCR

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