• Journal of Physiology & Pathology in Korean Medicine
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• v.23 no.3
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• pp.574-580
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• 2009

Melanogenesis is induced mainly by ultraviolet radiation of sunlight and ${\alpha}$-Melanocyte stimulation hormone (${\alpha}$-MSH) which binds to a specific G protein coupled receptor. ${\alpha}$-MSH and cAMP-elevating agents are known to melanin syntheisis and dendrite outgrowth. The purpose of this study was to investigate the mechanism of melanogenesis inhibition in B16/F10 cells by ethanol extract of Artemisiae Capillaris Herba. In the present study, ${\alpha}$-MSH led to a stimulation of melanin synthesis that appeared to result from an increased tyrosinase activity and melanin content. However, the ethanol extract of Artemisiae Capillaris Herba inhibited the ${\alpha}$-MSH-induced tyrosinase activity and melanin content. In control conditions, B16/F10 cells displayed a fibroblastic appearance while ${\alpha}$-MSH treatment promoted the emergence of small and numerous dendrites from the plasma membrane. The ethanol extract of Artemisiae Capillaris Herba abolished the ${\alpha}$-MSH-induced dendricity. Regarding protein levels of the melanogenic enzymes, the amounts of tyrosinase were increased after incubation with ${\alpha}$-MSH. The treatment of Artemisiae Capillaris Herba ethanol extract decreased the ${\alpha}$-MSH expression levels of tyrosinase. Based on these findings, it is likely that the ethanol extract of Artemisiae Capillaris Herba exerts its depigmenting effects in B16/F10 cells through the suppression of tyrosinase expression, which are key enzymes for melanogenesis.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.41 no.4
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• pp.383-389
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• 2015

In this study, we investigated the inhibitory effect on melanin synthesis of Ginkgo biloba seed oil. The results showed 9.96% inhibitory effect scavenging activity on DPPH and showed a value of 1.33 mM of $FeSO_4$ at a concentration of 0.06% in DMSO by using FRAP assay. G. biloba seed oil inhibited tyrosinase activity up tp 37.72% and suppressed the biosynthesis melanin up to 48.02% at 0.06% in B16/F10 mouse melanoma cell. In G. biloba seed oil treated group tyrosinase, TRP-1, TRP-2 and MITF gen expression levels significantly decreased compared to the contral group at a concentration of 0.04% and 0.06%. In conclusion, these results indicated that G. biloba seed oil extract have a good antimelanogenetic effects.

• The Korea Journal of Herbology
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• v.28 no.3
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• pp.69-74
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• 2013

Objectives : Since hypopigmentation is known to increase the risk of skin cancer, melanogenesis in the skin needs to be regulated. Here, we evaluated the melanogenesis stimulatory effects of a modified Goagium herbal remedy (HR) and HR+ox bile (Bos taurus domesticus) extract (OBE) to address hypopigmentation disorders. Methods : B16F10 melanoma cells were treated with different dosages of HR and HR+OBE for 24 to 48 h after 1 h of 10 nM ${\alpha}$-melanocyte stimulating hormone (${\alpha}$-MSH). After the treatment, cell viability, tyrosinase activity, melanin synthesis and the expression of genes related to melanin synthesis were measured and the regulation of the ${\alpha}$-MSH signalling through cAMP responding element binding protein (CREB) was determined. Results : HR and HR+OBE with the ranges of $15{\sim}100{\mu}g/mL$ did not affect cell viability in melanoma cells. The 1 h treatment of HR+OBE (50 and $100{\mu}g/mL$) potentiated the phosphorylation of CREB by enhancing ${\alpha}$-MSH signaling and its 24 h treatment increased CREB expression. Consistent with CREB potentiation, their treatment for 24 h, the expression of microphthalmia-associated transcription factor (MIFT), tyrosinase, tyrosinase related protein (TRP)-1 and TRP-2 were increased in realtime PCR. Ultimately, the 48 h treatment of HR+OBE (50 and $100{\mu}g/mL$) increased tyrosniase activity and melanin contents in the melanoma cells in comparison to the control. Conclusions : HR+OBE (50 and $100{\mu}g/mL$) increases melanin synthesis in B16F10 melanoma cells via the stimulation of tyrosinase activity and expression of MIFT, tyrosinase, TRP-1 and TRP-2. HR+OBE can be used as the a possible treatment for hypopigmentation of the skin.

• Proceedings of the KAIS Fall Conference
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• 2012.05a
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• pp.182-186
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• 2012

human tyrosinase (hTyr) catalyzes first and the rate limiting step in the synthesis of polymerized pigment, melanin which determines skin, hair and eye colors. Mutation of hTyr often brings about decrease of melanin production and further albinism. Meanwhile, a number of cosmetic companies providing skincare products for woman in Asia-Pacific region have tried to develop inhibitors to bright skin color for several decades. In this study, we built a 3D structure by comparative modeling technique based on the crystal structure of tyrosinase from bacillus megaterium as a template to serve structural information of hTyr. According to our model and sequence analysis of type 3 copper protein family proteins, two copper atoms of active site located deep inside are coordinated with six strictly conserved histidine residues coming from four-helix-bundle. Cavity which accommodates substrates was like funnel shape of which entrance was wide and expose to solvent. In addition, protein-substrate and protein-inhibitor complex were modeled with the guide of van der waals surface generated by in house software. Our model suggested that only phenol group or its analogs can fill the binding site near nuclear copper center because inside of binding site has narrow shape relatively. In conclusion, the results of this study may provide helpful information for designing and screening new anti-melanogensis agents.

• BMB Reports
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• v.42 no.3
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• pp.178-183
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• 2009

Tyrosinase (TYR) plays a critical role in cellular melanogenesis and, thus, has been the major target of pharmacological approaches for the control of skin pigmentation. This study examined an alternative molecular approach using TYR-small interfering RNA (siRNA) to control melanogenesis in the human melanocytes. Both the mRNA and protein levels of TYR were significantly lowered by TYR-siRNA treatment, whereas TYR-related protein 1 and TYR-related protein 2 displayed no such changes. TYR-siRNA treatment inhibited the cellular melanin synthesis from the externally supplied TYR substrate L-tyrosine. TYR-siRNA also suppressed melanin synthesis and decreased the viability of cells exposed to ultraviolet radiation, supporting a critical role of melanin in protection against ultraviolet radiation. These results suggest that molecular approaches using siRNA targeted to the enzymes of melanogenic pathway may provide a novel strategy for the control of cell pigmentation.

• Journal of Life Science
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• v.29 no.11
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• pp.1192-1199
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• 2019

The intention of this study was to confirm the possible use of an ethanol extracts of Nelumbinis Rhizomatis Nodus (NRN) as a cosmetic material. To this end, we extracted NRN with 70% ethanol and performed biological activity evaluation of whitening efficacy and wrinkle reduction. We performed cellular tyrosinase inhibition and melanin contents assay to check the whitening activity of NRN and carried out a toxicity evaluation of NRN via an MTT assay and the amounts of associated proteins that affect melanin production in a melanoma cell line (B16F10). And collagenase inhibitory assay was performed for the evaluation of anti-wrinkle of samples. In addition, a toxicity evaluation using an MTT assay and matrix metalloprotease (MMP-1) and procollagen synthesis inhibition by NRN were evaluated in a fibroblast cell line (CCD-986sk). Western blot results for the whitening activity evaluation revealed that the levels of two proteins related to melanin production, tyrosinase-related protein-1 (TRP-1) and tyrosinase-related protein-2 (TRP-2), were decreased in a dose-dependent manner. Moreover, collagenase inhibition activity at a concentration of $500{\mu}g/ml$ NRN by measuring epigallocatechin-3-gallate (EGCG) was increased by more than 80% compared to the control group. Meanwhile, procollagen synthesis was reduced by 68.8% in the UVB-induced CCD- 986sk cells group whereas collagen synthesis recovered by 80.2% with $25{\mu}g/ml$ NRN. The MMP-1 expression rate showed 20.2% reduction at $25{\mu}g/ml$. The results of the experiments verified the whitening and wrinkle suppression effects of NRN and confirmed that it could be used as a safe natural cosmetic material in the future.

• Applied Chemistry for Engineering
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• v.25 no.5
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• pp.468-473
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• 2014

Dendropanax morbifera (D. morbifera) grows in the southern coastal areas and on Jeju Island in Korea. In this study, D. morbifera leaf extract was investigated to determine the mechanism of its whitening effect. The inhibitory activities of the extract on melanogenesis were tested in B16 melanoma cells treated with the ${\alpha}$-melanocyte stimulating hormone (${\alpha}$-MSH). D. morbifera leaf extracts remarkably decreased the melanin content at 25 and $50{\mu}g/mL$. The extracts significantly inhibited the intracellular tyrosinase activity and protein expression of tyrosinase and tyrosinase related protein-2 (TRP-2). In conclusion, D. morbifera leaf extracts would show a whitening effect by inhibiting intracellular tyrosinase activities and the expression of enzymes directly involved in the melanin biosynthesis. The results indicate that fractions of D. morbifera leaf extracts show potential for application as a whitening agent in the new whitening cosmetics.

• Journal of Life Science
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• v.20 no.11
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• pp.1617-1624
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• 2010

Recently, it has been found that Asiasari radix showed a hypopigmenting effect on melanogenesis through activation of mitogen-activated protein kinase (MEK)/extracellular signal-activated kinase (ERK) in B16F10 melanoma cells. However, the hypopigmenting effect of A. radix on the $\alpha$-melanocyte stimulating hormone ($\alpha$-MSH)-stimulated melanogenesis has remained unknown. The purpose of this study was to investigate the inhibitory mechanism of the partially purified A. radix (PPAR)-induced hypopigmentating effects on $\alpha$-MSH-stimulated melanogenesis in B16F10 mouse melanoma cells. PPAR strongly inhibited tyrosinase activity and leads to decreased melanin synthesis in $\alpha$-MSH-stimulated B16F10 melanoma cells. PPAR also decreased the $\alpha$-MSH-induced over-expression of the melanogenic enzymes, tyrosinase, tyrosinase-related protein (TRP)-1, dopachrome tautomerase (Dct) and microphthalmia-associated transcription factor (MITF). We further showed that PPAR inhibits $\alpha$-MSH-induced melanogenesis via phosphorylation of MEK/ERK and PI3K/Akt, and that their activation was blocked by MEK inhibitors, PD98059 and PI3K inhibitors, LY294002 in $\alpha$-MSH-stimulated B16F10 melanoma cells. These results suggest that PPAR inhibits $\alpha$-MSH-induced melanogenesis by activation of MEK/ERK and PI3K/Akt through MITF degradation, which may lead to down-regulation of tyrosinase.

• ALGAE
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• v.30 no.2
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• pp.163-170
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• 2015

Tyrosinase inhibitors are an important component of cosmetic products. Our previous studies have proposed that eckol isolated from the brown alga Ecklonia cava, can be explored as a tyrosinase inhibitor. However, cellular activities and mechanism of action of eckol remain unknown. Therefore, the current study analyzed the eckol binding modes using the crystal structure of Bacillus megaterium tyrosinase. The effects of eckol on melanin synthesis induced by ${\alpha}$-melanocyte stimulating hormone in B16F10 melanoma cells were also investigated. We predicted the 3D structure of tyrosinase and used a docking algorithm to simulate binding between tyrosinase and eckol. These molecular modeling studies were successful (calculated binding energy value, $-115.84kcal\;mol^{-1}$) and indicated that eckol interacts with Asn205, His208, and Arg209. Furthermore, eckol markedly inhibited tyrosinase activity and melanin synthesis in B16F10 melanoma cells. We also found that eckol decreased the expression of tyrosinase, tyrosinase-related protein (TRP) 1, and TRP2. These results indicate that eckol is a potent inhibitor of melanogenesis, and this finding may be useful for the development of novel pharmaceutical and cosmetic agents.

• Journal of Life Science
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• v.22 no.3
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• pp.318-323
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• 2012

This study involved observation of the inhibitory effect of 70% EtOH and water extracts from Doinseunggitang on melanin synthesis, tyrosinase activity, and western blotting using B16F10 melanoma cells. Doinseunggitang extracts inhibited melanin synthesis and tyrosinase activity in a dependent manner. As a result, it was found that Doinseunggitang 70% EtOH extracts inhibit melanin synthesis and tyrosinase activity, respectively, by 40% and 51%. In addition, western blotting analysis showed that 70% EtOH extracts inhibited tyrosinase, MITF, TRP-1, and TRP-2 expression. These results show that 70% ethanol extracts of Doinseunggitang could be developed as a skin whitening material in cosmetics.