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PAPER REVIEW!

INHA University - 김준섭

Sulforaphane Inhibited Melanin Synthesis by Regulating Tyrosinase Gene Expression in B16 Mouse Melanoma Cells

▶Objective : Sulforaphane 과 Tyrosinase 와 Melanogenesis 에 대한 관계를 알아보기

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Sulforaphane is a compound widely present in consumed vegetables, par-ticularly broccoli. Previous studies have demonstrated that sulforaphane has many physiological effects including anti-cancer, anti-oxidation, and detoxification. In this study, we found that sulforaphane inhibited melano-genesis and tyrosinase expression. The inhibitory effect of 5M sulforaphane on melanogenesis was determined to be equivalent to that of 100M arbutin. In addition, sulforaphane induced phosphorylated extracellular signal-regu-lated kinase (ERK) and inhibited phosphorylated p38. It has been reported that the phosphorylated mitogenactivated protein (MAP) kinase family (ERK and p38) controls tyrosinase expression. Our data indicate that sul-foraphane inhibited melanogenesis and tyrosinase expression by affecting the phosphorylated MAP kinase family. These findings indicate that sul-foraphane might be an effective skin-whitening agent.

ABSTRACTS

5M sulforaphane 100M arbutin ≒

▶sulforaphane inhibits melanogenesis and tyrosinase expression.

▶MAP kinase : mitogen-activated protein kinase

▶ERK : extracellular signal-regulated kinase

sulforaphane<activation> phosphorylated ERK

<inhibition> phosphorylated p38

tyrosinase expression melanin

ERK p38

MAP kinase family

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<regulation>

SULFORAPHANE

sulforaphane

in vegetable, broccoli, broccoli sprouts

myrosinase : glucoraphanin sulforaphane

isothiocyanate group

skin-whitening agent

antioxidant, help remove ROS

exhibit anti-cancer and antimicrobial properties

< 출처 : wikipedia>4

INTRODUCTION

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▶In this study, we examined the inhibitory effect of sulforaphane on tyrosi-nase

▶key enzymes in melanogensistyrosinase, tyrosinase-related protein 1 (TRP-1), tyrosinase-related protein 2 (TRP-2)

▶MAP kinase family controls melanogenesis.

<induction> phosphorylated ERK

<inhibition> phosphorylated p38

MITF

melanin synthesis

▶skin is exposed to UV

1) αMSH

2) αMSH + MC1R(receptor)

3) MITF

4) tyrosinase expression

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MATERILALS AND METH-ODS

Cell culture : B16 murine melanoma cells

Measurement of melanin contents

Cell viability assay : electron microscope

Tyrosinase activity assay : Bradford assay, microplate

reader

Western blotting analysis

Statistical analysis : Dunnett’s test

RESULTS – MELANIN ASSAY

Fig. 1. Effects of Sulforaphane on Melanin Production in B16 Melanoma Cells.

Effects of sulforaphane on melanogenesis using B16 melanoma cells

Fig. 2. Cytotoxic Effects of Sulforaphane on B16 Melanoma Cells.

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“melanin 합성의 저해 !!”

“no cytotoxic effects”

RESULTS – TYROSINASE AS-SAY

Fig. 3. Effects of Sulforaphane on Tyrosi-nase Activity in B16 Melanoma Cells.

Effects of sulforaphane on tyrosinase expression in B16 melanoma cells

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▶in vitro tyrosinae assay tyrosinase 의 활성에 영향을 주지 않았다 .

▶cellular tyrosinase assay tyrosinase activity 가 감소

tyrosinase 의 수량이 감소 !

RESULTS – WESTERN BLOTTING AS-SAY

Fig. 4. Effects of Sulforaphane on Tyrosinase Expression in B16 Melanoma Cells.

Effects of sulforaphane on tyrosinase expression in B16 melanoma cells

9Fig. 5. Effects of Sulforaphane on MAP Kinase Pathways in B16 Melanoma Cells.

Effects of sulforaphane on MAP kinase activation in B16 melanoma cells

sulforaphane

tyrosinase

p-p38 (360-720

min)

p-ERK (60-720

min)

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DISCUSS Sulforaphane treatment inhibited p38 phosphorylation in B16 cells.

1. inhibition of p38 phosphorylation

induction of ERK phosphorylation

2. inhibition of MITF activation

3. inhibition of TRY, TRP1, TRP2 expres-

sion

4. inhibition of melanin synthesis

Sulforaphane