[논문]B16F10세포에서 멜라닌 생성 억제제 및 항산화제로서 조구등의 억제 효과

동원원; 우영민; 차지현; 차재영; 려내유; 백민우; 박준성; 이상현; 하종명; 김안드레

doi:10.5352/jls.2020.30.12.1033

B16F10세포에서 멜라닌 생성 억제제 및 항산화제로서 조구등의 억제 효과
The Effect of Inhibition of Uncaria rhynchophylla as an Inhibitor of Melanogenesis and an Antioxidant in B16F10 Melanoma Cells 원문보기

생명과학회지 = Journal of life science, v.30 no.12, 2020년, pp.1033 - 1041

동원원 (신라대학교 의생명과학 제약공학과) , 우영민 (신라대학교 자연과학연구소) , 차지현 (신라대학교 의생명과학 제약공학과) , 차재영 (신라대학교 의생명과학 제약공학과) , 려내유 (신라대학교 의생명과학 제약공학과) , 백민우 (신라대학교 의생명과학 제약공학과) , 박준성 (신라대학교 웰빙체육학부) , 이상현 (신라대학교 의생명과학 제약공학과) , 하종명 (신라대학교 의생명과학 제약공학과) , 김안드레 (신라대학교 의생명과학 제약공학과)

초록
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최근 모든 연령대의 사람들은 미용적인 이유로 더 밝은 피부를 원하고 있으며, 천연 제품은 화학적으로 합성된 화합물보다 더 많은 관심을 받고 있다. 조구등은 아시아에서 전통 한약재로 널리 사용되어 왔다. 새로운 피부 미백제를 찾기 위해 본 연구에서는 조구등의 항산화 활성과 잠재적인 tyrosinase 억제 작용을 확인하였다. 조구등을 70% 에탄올로 추출하여 항산화 활성을 분석하고 tyrosinase 활성과 melanin 합성에 미치는 영향을 평가했다. 총 mRNA 발현은 RT-PCR을 사용하였다. 그 결과 조구등 추출물은 B16F10 세포에서 뛰어난 항산화 능력과 상당한 수준의 폴리페놀 및 플라보노이드 화합물을 함유하였다. 또한, 세포 내 tyrosinase 활성을 억제하고 처리된 세포에서 melanin의 양을 감소시켰다. Tyrosinase의 mRNA 발현은 1 mg/ml 농도에서 현저히 감소하였다. 이와 같은 결과는 조구등이 미백 효과가 있는 화장품의 천연 소재로 사용될 수 있는 높은 잠재력을 가지고 있음을 시사한다.

Abstract ▼ AI-Helper

Many people of all ages wish to have lighter skin for cosmetic reasons, and natural products attract more attention than chemically synthesized compounds. Uncaria rhynchophylla is widely used in Asia as a traditional herbal medicine. In order to find novel skin whitening agents, the present study evaluated the antioxidant activity and potential tyrosinase-inhibiting properties of U. rhynchophylla. Specifically, this study analyzed the antioxidant capacity of a 70% ethanolic extract of U. rhynchophylla as well as its effects on tyrosinase activity and melanin synthesis. Total mRNA levels were examined using reverse transcription polymerase chain reaction. The results revealed that U. rhynchophylla extracts exhibit great antioxidant capacity and significant levels of polyphenol and flavonoid compounds. U. rhynchophylla extracts can also powerfully inhibit tyrosinase activity. This same capacity was observed in melanoma B16F10 cells; that is, U. rhynchophylla extracts suppressed intracellular tyrosinase activity and reduced the amount of melanin in treated cells. In addition, a 1 mg/ml concentration of U. rhynchophylla extract significantly reduced the mRNA expression levels of tyrosinase. U. rhynchophylla extracts decrease tyrosinase and inhibit melanogenesis in B16F10 cells. This finding suggests that U. rhynchophylla has great potential as a natural whitening agent in skincare products.

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표/그림 (7)

그림 Fig. 1. The antioxidant properties of U. rhynchophylla. (A) DPPH free radical scavenging activity; (B) ABTS radical scavenging capacity. Various concentrations of the U. rhynchophylla extracts and vitamin C were as a positive standard in the above assays. URE, 70% ethanolic extract of U. rhynchophylla. All values are expressed as mean ± S.D. (n=3). The letters a, b, c, d, and e above the bars stand for significantly different (p<0.05) groups by one-way ANOVA, followed by Duncan's multiple test.
그림 Fig. 2. Polyphenol and flavonoids contents of U. rhynchophylla. (A) Measurement of polyphenol contents; (B) Evaluation of flavonoid contents. URE, 70% ethanolic extract of U. rhynchophylla. All values are expressed as mean ± S.D. (n=3). The letters a, b, c, and d above the bars stand for significantly different (p<0.05) groups by one-way ANOVA, followed by Duncan's multiple test.
그림 Fig. 3. Effect of U. rhynchophylla on mushroom tyrosinase activ-ity. URE, 70% ethanolic extract of U. rhynchophylla. All values are expressed as mean ± S.D. (n=3). The letters a, b, and c above the bars stand for significantly different (p<0.05) groups by one-way ANOVA, followed by Dun-can's multiple test.
그림 Fig. 4. Effect of U. rhynchophylla on cell viability in B16F10 cells. The cells were seeded at 6×10³ cells per well and in-cubated in media containing 0.1-1 mg/ml concentration of URE for 24 hr. URE, 70% ethanolic extract of U. rhynchophylla. All values are expressed as mean ± S.D. (n=3).
그림 Fig. 5. Effect of U. rhynchophylla on melanin content. (A) Cell morphology was observed under a phase contrast microscope. (B) Effect of U. rhynchophylla on melanin content in B16F10 cells. The cells were treated with 0.1-1 mg/ml of URE or kojic acid (positive control) for 24 hr after treatment with/without 200 nM α-MSH for 4 hr. URE, 70% ethanolic extract of U. rhynchophylla. All values are expressed as mean ± S.D. (n=3). The letters a, b, c, and d above the bars stand for significantly different (p<0.05) groups by one-way ANOVA, followed by Duncan's multiple test.
그림 Fig. 6. Effect of U. rhynchophylla on cellular tyrosinase activity. The cells were treated with 0.1-1 mg/ml of URE or kojic acid as positive control for 24 hr after treatment with/without 200 nM α-MSH for 4 hr. The supernatant lysed in RIPA buffer was incubated with 5 mM L-DOPA for 1 hr and each sample was normalized to total celluar protein. URE, 70% ethanolic extract of U. rhynchophylla. All values are expressed as mean ± S.D. (n=3). The let-ters a, b, c, d, and e above the bars stand for significantly different (p<0.05) groups by one-way ANOVA, followed by Duncan's multiple test.
그림 Fig. 7. Effect of U. rhynchophylla on the mRNA expression of tyrosinase. Total RNA from B16F10 cells treated with U. rhynchophylla after 200 nM α-MSH collected at the indicated time. URE, 70% ethanolic extract of U. rhyncho-phylla. Total mRNA levels were examined by RT-PCR, using glyceraldehyde 3-phosphate dehydrogenase (GADPH) as an internal control. The graph represents the fold val-ue of exposed band intensity.

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대상 데이터

Kojic acid, L-DOPA, tyrosine, 3-(4, 5-dimethyl-2-thiazolyl)- 2, 5-diphenyl-tetrazolium bromide (MTT), diphenyl-1-picrylhydrazyl (DPPH), 2, 2’-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid (ABTS)), and dimethyl sulfoxide (DMSO) were purchased from Sigma-Aldrich (St Louis, MO, U.S.).

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