학술논문
B16F10 마우스 흑색종 세포에서 Tussilagone의 멜라닌 생성 억제 효과 / The Effects of Tussilagone on Anti-Melanogenesis in B16F10
Document Type
Dissertation/ Thesis
Author
Source
Subject
Language
Korean
Abstract
Tussilagone은 암에서의 항암, 항염증 등에 관해 밝혀져 있다. 하지만 피부 미백분야에서는 아직 알려지지 않아 B16F10 마우스 흑색종 세포주를 사용하여 Tussilagone의 멜라닌 합성 억제 효과를 밝히기 위해 세포 생존율, 멜라닌 분비 및 함량분석, Tyrosinase 억제, Tyrosinase 활성, mRNA(messenger ribonucleic acid)발현, 단백질 발현을 측정하였다. α-MSH 처리된 B16F10 세포에서Tussilagone은 멜라닌 분비와 멜라닌 생성 함량을 억제하였으며, 세포 외 Tyrosinse억제를 증가, 세포 내 Tyrosinase 활성을 감소시켜 멜라닌 생성을 감소시키는 것으로 밝혀졌다. 또한, microphthalmia-associated transcription factor(MITF), Tyrosinase-related protein 1(TYRP1), Tyrosinase-related protein2(TYRP2), Tyrosinase(TYR)의 mRNA발현 수준에서 확인한 결과, Quantitative real-timePCR(q-PCR)에서 TRP1을 제외한MITF/TYRP2/TYR의 발현 수준을 감소시키는 것으로 나타났다. mRNA수준에서 MITF를 Tussilagone최고 농도에서 발현을 감소시키는 것을 확인하여 멜라닌 합성 관련 단백질 수준에서 확인한 결과, Tussilagone의 높은 농도인 20, 25μM에서 p-CREB의 발현을 감소되어 멜라닌 생성을 억제하는 것을 확인하였다. 이 데이터들은 Tussilagone이 B16F10마우스 흑색종 세포에서 멜라닌 합성을 억제하여 미백 효과를 증가시킨다는 것을 나타냈다. 따라서 Tussilagone은 피부 미백에 효과적인 성분임을 나타내며, 기능성 물질을 개발하기 위한Tussilagone의 작용기전에 대해 추가적인 연구가 필요하다.
This study was conducted to elucidate the inhibitory effect of Tussilagone on melanin synthesis. In order to elucidate the effects of Tussilagone on the B16F10 mouse melanoma cell line, cell viability, melanin production, Tyrosinase inhibition, Tyrosinase activity assay, and messenger ribonucleic acid (mRNA) expressions were measured. In α-MSH-treated B16F10 cells, Tussilagone suppressed melanin secretion and melanogenesis, increased extracellular Tyrosinase inhibition, and decreased intracellular Tyrosinase activity, thereby reducing melanogenesis. In addition, determination of the mRNA expression levels of microphthalmia- associated transcription factor (MITF), Tyrosinase-related protein 1 (TYRP1), Tyrosinase- related protein 2 (TYRP2), and Tyrosinase (TYR) by quantitative real-time PCR (q-PCR) revealed a decrease in the expression levels of MITF/TYRP2/TYR but not TRP1. It was confirmed that expression of MITF at the mRNA level was decreased at the highest concentration of Tussilagone. Proteins related to melanin production were quantified at the protein level, confirming that the expression of p-CREB was reduced at high concentrations of 20 and 25 μM of Tussilagone, thereby suppressing melanin production. These data indicate that Tussilagone increased the whitening effect by inhibiting melanin synthesis in B16F10 mouse melanoma cells. Thus, Tussilagone appears to be an effective ingredient for skin whitening, and further research on the mechanism of action of Tussilagone for the development of functional materials should be investigated.
This study was conducted to elucidate the inhibitory effect of Tussilagone on melanin synthesis. In order to elucidate the effects of Tussilagone on the B16F10 mouse melanoma cell line, cell viability, melanin production, Tyrosinase inhibition, Tyrosinase activity assay, and messenger ribonucleic acid (mRNA) expressions were measured. In α-MSH-treated B16F10 cells, Tussilagone suppressed melanin secretion and melanogenesis, increased extracellular Tyrosinase inhibition, and decreased intracellular Tyrosinase activity, thereby reducing melanogenesis. In addition, determination of the mRNA expression levels of microphthalmia- associated transcription factor (MITF), Tyrosinase-related protein 1 (TYRP1), Tyrosinase- related protein 2 (TYRP2), and Tyrosinase (TYR) by quantitative real-time PCR (q-PCR) revealed a decrease in the expression levels of MITF/TYRP2/TYR but not TRP1. It was confirmed that expression of MITF at the mRNA level was decreased at the highest concentration of Tussilagone. Proteins related to melanin production were quantified at the protein level, confirming that the expression of p-CREB was reduced at high concentrations of 20 and 25 μM of Tussilagone, thereby suppressing melanin production. These data indicate that Tussilagone increased the whitening effect by inhibiting melanin synthesis in B16F10 mouse melanoma cells. Thus, Tussilagone appears to be an effective ingredient for skin whitening, and further research on the mechanism of action of Tussilagone for the development of functional materials should be investigated.