• Molecular & Cellular Toxicology
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• v.2 no.2
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• pp.81-88
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• 2006

Sphingolipid metabolites regulate many aspects of cell proliferation, differentiation, and apoptosis. In the present study, we have assessed the effects of the novel phytosphingosine derivative, N-acetylphytospingosine (NAPS), on the depigmentation of murine B16F10 melanoma cells, and have also attempted to identify the possible signaling pathway involved, in comparison with $C_{2}-ceramide$. NAPS and $C_{2}-ceramide$ both inhibited the growth of the B16F10 cells in a dose-dependent manner. Melanin content and tyrosinase activity were significantly reduced in response to treatment with NAPS and $C_{2}-ceramide$ at concentrations in a range between $1-5\;{\mu}M$. However, the levels of tyrosinase mRNA, as well as the levels of tyrosinase related protein-1 (TRP-1) and tyrosinase related protein-2 (TRP-2) genes and the level of tyrosinase protein remained unaffected by treatment with either NAPS or $C_{2}-ceramide$. We also attempted to determine the signaling pathway exploited by NAPS and $C_{2}-ceramide$. Interestingly, the phosphorylation of Akt/PKB at serine 473 by NAPS was reduced at the 5 minute mark, whereas $C_{2}-ceramide$ induced the phosphorylation of Akt/PKB at serine 473. Finally, Akt/PKB activity in the NAPS-treated cells was elevated in comparison with the untreated cells. LY294002, a specific PI3-K inhibitor which is located upstream of Akt/PKB, inhibited the phosphorylation of Akt/PKB, but induced an increase in melanin synthesis. These results suggest that the activation of Akt/PKB at serine 473 is related with the suppression of melanin production in the B16F10 mouse melanoma cells. Therefore, the mechanisms exploited by NAPS and $C_{2}-ceramide$ responsible for the depigmentation of B16F10 cells were concluded to involve the inhibition of melanosomal tyrosinase activity.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.28 no.2
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• pp.1-12
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• 2015

Objective : The goal of this study is to identify the effects of extract ofGentianae sino-ornata(GSO) on the anti-oxidative activity of skin.For this purpose, several functions of GSO were analyzed in terms of skin-lightening activity and wrinkle improvement. Methods : Cell viability was measured by neutral red (NR) assay, and GSO showed highly efficacy in DPPH radical scavenging activity. The level of tyrosinase and matrix metalloproteinase-1 (MMP-1) in media was analyzed by ELISA kit, and the expressions of p-JNK and p-ERK was measured by Western blot. To elucidate inhibitory effects of GSO on melanin synthesis, I determined the tyrosinase activity and melanin production in B16F10 cells. Results : MMP-1 production in UVB-stimulated HDF cells was inhibited by GSO treatments, and also GSO inhibited protein expression levels of p-JNK and p-ERK. GSO significantly reduced tyrosinase activity and melanin synthesis in B16F10 cells. Conclusions : From these results, GSO appears to be effective on skin elasticity increase, wrinkle improvement, whitening as anti-aging activity.

• Journal of Physiology & Pathology in Korean Medicine
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• v.25 no.3
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• pp.510-515
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• 2011

This study was conducted to evaluate the depigmenting properties of ethanol extract from a Fritillaria verticillata Willd. (EFV) in B16F10 cells. Fritillaria verticillata Willd., a perennial herbaceous plant, has been used as a stimulator of mammary gland, expectorant, blood pressure depressant, antitussive agents in Korean herbal medicine. In the present study, we observed that melanin synthesis of B16F10 cells were significantly decreased by EFV without cytotoxicity. However, EFV could not suppress tyrosinase activity in B16F10 cells and mushroom tyrosinase activity. Furthermore, EFV did not effect the protein expression of tyrosinase, tyrosinase-related protein -1 (TRP-1), and TRP-2. These results suggest that EFV inhibited melanin synthesis and the hypopigmentary effect of EVF was not due to regulation of tyrosinase protein.

• Journal of Physiology & Pathology in Korean Medicine
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• v.21 no.6
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• pp.1456-1461
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• 2007

Melanogenesis is induced mainly by ultraviolet radiation of sunlight and ${\alpha}-melanocyte$-stimulating hormone (${\alpha}-MSH$) which binds to a specific G protein coupled receptor. The purpose of this study was to investigate the mechanism of melanogenesis inhibition in B16/F10 cells by methanol extract of Rubus coreanus Miquel (RCM). In the present study, ${\alpha}-MSH$ and forskolin led to a stimulation of melanin synthesis that appeared to result from an increased tyrosinase activity and melanin content. However, RCM inhibited the ${\alpha}-MSH$- and forskolin-induced melanin synthesis. In addition, RCM abolished the ${\alpha}-MSH$- and forskolin-induced cytoplasmic dendricity. Regarding protein levels of the melanogenic enzymes, the amounts of tyrosinase and tyrosinase-related protein 1 (TRP-1) were increased after incubation with α-MSH and forskolin. The treatment of RCM decreased the ${\alpha}-MSH$- and forskolin-induced expression levels of tyrosinase and TRP-1. Based on these findings, it is likely that RCM exerts its depigmenting effects in B16/F10 cells through the suppression of tyrosinase and TRP-1 expression, which are key enzymes for melanogenesis.

• Korean Journal of Pharmacognosy
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• v.45 no.3
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• pp.262-267
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• 2014

Royal jelly composes of many components, especially protein. Protein is a major factor which cause allergy. We focused on water soluble royal jelly (WSRJ) that was removed allergy - inducing protein. 10-hyroxy-2-decenoic acid content of WSRJ is 2.42 g/100 g, which is double compared to that of lypophilized RJ. To further access WSRJ as a cosmetic ingredient and potential external treatment for topical use, we investigated its ability to inhibit tyrosinase activity and melanin biosynthesis on melanogenesis in B16F1 melanoma cells. We found that WSRJ increased the cell viability in B16F1 melanoma cell and WSRJ (1~10 mg/ml) inhibited melanin synthesis in with 10 nM ${\alpha}$-melanocyte-stimulating hormone (${\alpha}$-MSH) for 48 h. WSRJ inhibited direct tyrosinase activity, which decreased melanin synthesis in ${\alpha}$-MSH stimulated B16F1 melanoma cells. Thease findings suggest that WSRJ induces the down regulation of melanogenesis by inhibiting tyrosinase activation.

• Journal of Physiology & Pathology in Korean Medicine
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• v.29 no.3
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• pp.223-229
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• 2015

In this study, we investigated the effect of ethanol extracts from Gentianae scabrae bunge (GSB) on the activities of antioxidant, whitening and anti-inflammation. Viability of cells was measured by neutral red (NR) assay, and inhibitory effects of GSB on melanin synthesis was determined the melanin production in B16F10 cells. The expression level of matrix metalloproteinase-1 (MMP-1) in media was analyzed by ELISA kit, and nitric oxide (NO) production in RAW264.7 cells was monitored by measuring the nitrite content in culture medium. GSB showed highly efficacy in 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity, and significantly reduced melanin synthesis in B16F10 cells. MMP-1 production in UVB-stimulated human dermal fibroblast (HDF) cells was inhibited by GSB treatments. NO production was suppressed by the treatment of GSB in LPS-stimulated RAW264.7 cells. From this results, it was indicated that GSB could be utilized as anti-aging and whitening cosmetic ingredients.

• Journal of Life Science
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• v.19 no.3
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• pp.384-389
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• 2009

This study was carried out to elucidate the relation between expression levels of three melanin synthesis genes (Tyrosinase, Tyrosinase-related protein 1 and Dopachrome tautomerase) according to the Melanocortin-1 receptor genotypes with coat color patterns in Hanwoo cattle, Jeju black cattle and Holsteins. Using real-time semiquantitative reverse transcription-PCR assay (RT-PCR), the expression levels of these three genes were analyzed in skin tissues from four representative coat colored areas: yellowish-brown from MC1R e/e Hanwoo, wild type black from $E^+/E^+$ Jeju black cattle (JBC), and dominant black and white pied regions from $E^D/E^D$ Holstein. The TYR, TYRP1 and DCT genes showed higher expression levels of 4.5, 2.3 and 2.5 times higher in the black skin area of Holsteins than those of from JBC, respectively (p<0.001). In addition, the expression levels of these three genes from JBC were significantly higher than those from Hanwoo cattle (p<0.001). These results show that coat color phenotypes in Hanwoo cattle, JBC and Holsteins is directly correlated with TRY, TYRP1 and DCT transcription levels, which probably reflected involvement with MC1R genotypes; e/e in Hanwoo, $E^+/E^+$ in JBC and $E^D/E^D$ in Holsteins. Consequently, this study suggested that the status of MC1R protein may not only induce the transcription activities of a series of TYR and its related genes responsible for melanin synthesis, but also determine the levels of total melanin contents in bovine skin.

• Journal of Sericultural and Entomological Science
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• v.51 no.2
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• pp.207-210
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• 2013

In this study, matured silkworm hemolymph was studied for inhibitory effect on melanin formation, tyrosinase activities. A matured silkworm hemolymph mass purification method was developed and some of suppressing melanin formation effects were investigated, too. As a result, the advanced purification method was higher in yield rate than existing one by 53%. A free radical scavenging activity of hemolymph was higher than sericin on the market($IC_{50}$, $202.9{\pm}33.1$) and lower than vitamin C($0.57{\pm}0.02$) by $6.4{\pm}0.3{\mu}g/m{\ell}$. An inhibitory effect on melanin synthesis was higher in hemolymph compared to Morus bark and arbutin by 9.15% and 11.56%, respectively. An inhibitory effect on tyrosinase activity was higher in hemolymph than Morus bark and lower than arbutin. Effect of hemolymph $45{\mu}g/m{\ell}$ on suppressing of tyrosinase was almost 90%. Thus, these results suggest that hemolymph of matured silkworm may have beneficial properties as a material for cosmetics.

• Journal of Applied Biological Chemistry
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• v.65 no.3
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• pp.231-237
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• 2022

Biorenovation is a method that converts existing compounds into new compounds through the enzymatic action of microorganisms. Biorenovation has expected effects such as reducing toxicity of compounds and increasing their activity. In this study, we successfully synthesized Luteolin-7-O-glucoside (L7G) through biorenovation and investigated its inhibitory effect on melanin production in α-Melanocyte stimulating hormone induced B16F10 mouse melanoma cells. We confirmed that Luteolin was toxic at 50, 100 and 200 µM, but our L7G in same concentration was not toxic for B16F10 mouse melanoma cells and also showed significant reduction in melanin production and tyrosinase activity. In addition, while investigating the effect of L7G on factors involved in melanin synthesis through western blotting, we were able to confirm that the MITF and tyrosinase protein synthesis was inhibited in treatment with L7G, however, tyrosinase related protein-1 (TRP-1) and dopachrome tautomerase (TRP-2) expression was not affected. So we derived a conclusion that through biorenovation we could produce compounds like L7G with improved activity and reduced toxicity for possible use as an active ingredient with whitening functionality in cosmetics.It also suggests that the application of biorenovation has potential usefulness in developing anti-inflammatory materials. It also suggests that the application of bio-renovation has potential usefulness in the development of inflammatory material. We applied Biorenovation technology to Distylium racemosum extract (DR) to generate Distylium racemosum biorenovation product (DRB), and investigated the anti-inflammatory properties of DRB in lipopolysaccharide (LPS)-treated RAW264.7 macrophages. We are applying technology to Biorenovation Distylium racemosum extract (DR) Distylium racemosum was to create a biorenovation product (DRB), lipopolysaccharide (LPS) investigated the anti-inflammatory properties of DRB in RAW264.7 macrophages treated for.

• Proceedings of the SCSK Conference
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• 2003.09a
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• pp.763-764
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• 2003

As melanin is a key material for skin pigmentation, inhibitors of melanin formation have been used to cosmetics and drugs to prevent hyperpigmentation. Therefore, search for effective inhibitor$ from various plants were attempted. For this purpose, I examined in vitro tyrosinase assay system. Tyrosinase showed a maximal activity at 4 units concentration of tyrosinase, 10 minutes, 42$^{\circ}C$ and pH 6.5.