• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
• /
• v.28 no.1
• /
• pp.41-52
• /
• 2015

Objective : This study investigated the inhibitory mechanism of the hypopigmentating effects on ethanol extract of Rosae rugosae Flos (ERR) that has not yet been examined. Methods : We analyzed the anti-melanogenic effects of ethanol extracts from Rosae rugosae Flos by tyrosinase activity, melanin contents. We also examined protein expression levels of tyrosinase, TRP-1, TRP-2, MITF and ERK by western blot analysis in melanoma cells. Results : In this investigation, ERR effectively reduced ${\alpha}$-MSH-stimulated melanin synthesis by suppressing expression of tyrosinase and tyrosinase-related protein-1 (TRP-1). On the other hand, the expression of tyrosinase-related protein-2 (TRP-2) were not affected by treatment with ERR. ERR inhibited the expression of microphthalmia-associated transcription factor (MITF) as a key transcription factor for tyrosinase expression regulating melanogenesis. The upstream signaling pathway including cAMP response element-binding protein (CREB) and MAPKs were also inhibited by ERR. Pretreatment with PD98059, ERK inhibitor, attenuated the inhibitory effect of ERR on ${\alpha}$-MSH-induced tyrosinase activity. Conclusions : Our study suggested that the anti-melanogenic activity of ERR is correlated with the suppression of tyrosinase gene through CREB/MITF/ERK pathway.

• YAKHAK HOEJI
• /
• v.45 no.3
• /
• pp.269-275
• /
• 2001

The inhibitory effect of extract of Atractylodis Rhizoma alba on melanin biogenesis was studied by using B16/F10 melanoma in culture. Atractylodis Rhizoma alba significantly inhibited tyrosinase activity, and melanin contents with or without $\alpha$-MSH and forskolin in vitro. Melanin contents and tyro-sinase activity have decreased in a dose-dependent manner. These results show that extract of Atractylodis Rhizoma alba could be developed as skin whitening components of cosmetics.

• YAKHAK HOEJI
• /
• v.47 no.1
• /
• pp.31-36
• /
• 2003

To investigate the effect of protein kinase on melanin production via cAMP-dependent pathway, we measured the melanin amount and tyrosinase activity in B16 melanoma cells stimulated by alpha-melanocyte stimulating hormone (MSH), forskolin and 8-Br-cAMP. MSH, forskolin and 8-Br-cAMP significantly increased both melanin production and tyrosinase activity in B16 cells. Melanin production and tyrosinase activity by MSH are significantly inhibited by cyclic AMP-dependent protein kinase inhibitor (KT5720) and protein kinase C down-regulation treated with PMA. Bisindolmaleimide (1$\mu$M), protein kinase C inhibitor, significantly inhibited melanin production and tyrosinase activity stimulated by MSH, forskolin and 8-Br-cAMP with the following order of potency: MSH＞forskolin＞8-Br-cAMP. Tyrosine kinase inhibitor, genistein and DHC, significantly inhibited both, but the inhibitory effect was more potent in 8-Br-cAMP-stimulated B16 cells than MSH-stimulated cells. NFkB inhibitor (parthenolide) significantly inhibited melanin production and tyrosinase activity. Neither melanin production nor tyrosinase activity induced by MSH, forskolin and 8-Br-cAMP were affected by KN-62 (calmodulin-dependent protein kinase II inhibitor), PD098059 (mitogen-activated protein kinase inhibitor, MAPKK) and worthmannin (phosphatidylinositol 3-kinase inhibitor). These results suggest that both protein kinase C and tyrosine kinase are involved in melanin production by cyclic AMP-dependent pathway and NFkB pathway may play an important role in cyclic AMP-dependent melanin production in B16 melanoma cells.

• Korean Journal of Veterinary Research
• /
• v.58 no.2
• /
• pp.65-72
• /
• 2018

The present study observed the effects of a green tea (Camellia sinensis) flower extract (GTFE) on melanin synthesis in B16-F10 melanoma cells. GTFE exhibited antioxidant activity on 2,2-diphenyl-1-picrylhydrazyl and inhibited mushroom tyrosinase activity in a dose-dependent manner. Furthermore, GTFE significantly diminished ${\alpha}-melanocyte$ stimulating hormone (${\alpha}-MSH$) stimulated cellular melanin content and tyrosinase activity throughout the concentration range evaluated. Based on RNA sequencing analysis, differential gene expression patterns observed in ${\alpha}-MSH$ stimulated B16-F10 melanoma cells were normalized by the addition of GTFE. In particular, the expression levels of melanoregulin and tyrosinase genes which are key regulating genes in melanin synthesis were up-regulated by 3.5 and 3 fold respectively by ${\alpha}-MSH$, and were normalized to control levels by the addition of GTFE. The results suggest that GTFE inhibits melanin synthesis in ${\alpha}-MSH$ stimulated B16-F10 melanoma cells by normalizing expression of genes that are essential for melanin synthesis. Overall, the results suggest that GTFE could be applied in the development of a whitening agent for the treatment of dermal hyperpigmentation.

• Fisheries and Aquatic Sciences
• /
• v.22 no.5
• /
• pp.11.1-11.8
• /
• 2019

Background: In the present study, the skin-whitening effects of a marine-sourced mixture that includes a fucoidanrich extract of Undaria pinnatifida (UPEF), a phlorotannin-rich extract of Ecklonia cava (ECE), and glycosaminoglycans (GAGs) from sea squirt skin were investigated. Methods: The whitening effects of the mixture and its components were evaluated by measuring the inhibition of mushroom tyrosinase and melanin synthesis in alpha-melanocyte-stimulating hormone (${\alpha}$-MSH)-stimulated B16F10 melanoma cells. Results: Each component alone markedly inhibited mushroom tyrosinase in a dose-dependent manner, and in ${\alpha}$-MSH-stimulated B16F10 cells, they inhibited melanin synthesis and were cytotoxic. However, the whitening effects of UPEF, ECE, and GAGs in combination were greater than those of each component alone. A mixture in the ratio of 4:5:1 (UEG-451) showed the strongest activity without cytotoxicity. Further study suggested that UEG-451 inhibits ${\alpha}$-MSH-stimulated melanogenesis in B16F10 cells by downregulating tyrosinase and tyrosinase-related proteins, such as TRP-1 and TRP-2, via the inhibition of MITF expression. Conclusions: These results suggest that mixing the different components at optimum ratios might be an effective way to improve their bioactivities and reduce toxicity and that UEG-451 possesses strong whitening effects that could be used in the cosmetic industry.

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

• Korean Journal of Plant Resources
• /
• v.22 no.5
• /
• pp.477-482
• /
• 2009

Inhibitory effect of skin and seed of three species grape cultivated in Korea on melanogenesis was investigated. Melanin generation was examined in $\alpha$-Melanin Stimulating Hormone-stimulated B16 cell, mouse melanoma, in the presence of samples. All skin sample did not show the inhibitory effect. Seed extract of Campbell early and Neo Muscat had negative effect on cell viability. When $50{\mu}g/ml$ seed extract of Muscat Bailey A was treated, amount of generated melanin and cell viability were $51.6{\pm}20.5%$ and $90.4{\pm}11.3%$ compared to control, respectively. Seed extract of Muscat Bailey A reduced the tyrosinase protein induced by $\alpha$-Melanin Stimulating Hormone, which suggests that inhibitory effect of seed extract of Muscat Bailey A on melanin is partly due to suppression of tyrosinase that is responsible for melanin production.

• Journal of the Society of Cosmetic Scientists of Korea
• /
• v.24 no.3
• /
• pp.65-72
• /
• 1998

In this study, we demonstrated the whitening effect of Lagenaria leucantha through the melanin biosynthesis of S bikiniensis and inhibition of melanogenesis in cultured Bl6 melanocytes. And we confirmed the whitening effect of Lagenaria leucantha through the depigmentation of gold fish in vivo. The melanogenesis of B$_{16}$ melanocytes was founded to be activated dose and time dependently by the treatment of u- MSH. When the B$_{16}$ melanocytes was treated with 200nM of $\alpha$-MSH, the morphology of melanocytes was remarkably changed. The melanin content and the synthesis of tyrosinase were strikingly increased. Lagenaria ieucantha inhibited the melanin formation stimulated by $\alpha$-MSH without affection of cell viability. However, Lagenaria leucantha didn't inhibit tyrosinase activity and showed weak suppression on the synthesis of tyrosinase. These results suggest Lagenaria leucantha might inhibit melanin formation with tyrosinase independent manner. Lagenaria ieucantha also inhibition melanin biosynthesis with 18mm inhibition zone in S.bikiniensis. To evaluate the inhibitory activity of melanogenesis of Lagenaria leucantha in vivo, we examined its effect on depigmentation of gold fish. Lagenaria ieucantha remarkably reduced the size and density of melanophores in gold fish. These results suggest that Lagenaria ieucantha can be used as a whitening agent in cosmetics.ics.s.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
• /
• v.23 no.1
• /
• pp.149-157
• /
• 2010

목적 : 월견초는 다량의 불포화 지방산인 리놀렌산과 감마 리놀렌산을 함유하고 있으며, 천식성 기침이나 아토피피부염에 효능이 있는 것으로 알려져 있다. 본 연구는 월견초의 전초와 종자 추출물의 피부 멜라닌 합성에 대한 억제 효과를 조사하였다. 방법 : B16F10 멜라닌세포주를 이용하여 멜라닌, tyrosinase 활성 및 세포생존율을 측정하였다. 또한 멜라닌 합성- 관련효소인 tyrosinase, TRP-1, TRP-2의 단백질발현과 $\alpha$-MSH를 처리하여 색소침착을 유도 한 뒤 단백질 발현을 조사하였다. 결과 : 월견자는 B16F10 세포의 멜라닌 합성을 $5\;{\mu}g/ml$와 $10\;{\mu}g/ml$ 농도에서 각각 대조군의 81.3%, 68.3%로 억제하였고 tyrosinase의 활성도 이와 유사하게 억제하였다. 멜라닌 합성-관련효소들의 단백질발현을 관찰한 결과 월견초와 월견자는 tyrosinase 발현을 억제하였으며 TRP-1과 TRP-2의 발현에는 영향을 주지 않았다. 특히 $\alpha$-MSH에 의한 과색소 유도 시 tyrosinase 발현이 현저하게 감소되었으며, 월견자의 멜라닌 합성 억제 효과가 월견초 보다 높게 나타났다. 결론 : 이상의 연구 결과 월견자는 멜라닌세포의 tyrosinase 단백질 발현과 tyrosinase 활성을 억제하여 멜라닌 생성을 감소시키는 것으로 사료된다.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.20 no.5
• /
• pp.1271-1274
• /
• 2006

We investigated the effects of aqueous extracts from twelve medical herbs on ${\alpha}$-melanocyte stimulating hormone (${\alpha}$-MSH)-induced melanogenesis in B16F10 mouse melanoma cell. The cells were incubated with ${\alpha}$-MSH and aqueous extracts 5 days and 2 days and then analysed melanin amount and tyrosinase activities, respectively. Nine aqueous extract of herbs examined at 1 mg/m${\ell}$ level decreased melanin synthesis in B16F10 cell, especially in Agastache rugosa, Leonurus siviricus and Murus bombycis. The significant decrease of released extracellular melanin were also observed in treated cells with aqueous extract from Leonurus siviricus, Murus bombycis and Ledebouriella seseioides. The ${\alpha}$-MSH-induced activation of tyrosinase was inhibited in cells treated with aqueous extract from Cuscutae semen, Angelica tenuissima and Agastache rugosa. These results suggest that herbs inhibiting melanogenesis through tyrosinase activity may apply to develop whitening drugs and cosmetics.