• Journal of Applied Biological Chemistry
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• v.65 no.2
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• pp.93-99
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• 2022

Lysosome organelle extract (LOE) was derived from egg whites. The lysosome is an intracellular organelle that contains several hydrolysis enzymes. Previous studies have reported that LOE performs important functions, such as melanin de-colorization and anti-melanin production in B16F10 melanoma cells. However, its principal molecular and cellular mechanisms have not been elucidated till date. In non-cytotoxic conditions, LOE significantly inhibited α-MSH induced melanin synthesis of murine B16F10 cells. The anti-melanogenic activity of LOE was mediated by suppressing the mRNA expression of tyrosinase enzyme, tyrosinase related protein-1/2 (TRP-1/2), and microphthalmia-associated transcription factor (MITF) genes. By performing western blot analysis, we found that LOE significantly attenuated melanogenesis. In this case, LOE helped in increasing extracellular receptor kinase (ERK) phosphorylation in α-MSH induced B16F10 cells. Furthermore, MITF is found to be a key regulatory transcription factor in melanin synthesis; it was down-regulated by LOE through ERK phosphorylation. In this experiment, PD98059 (MEK inhibitor) was used to check whether LOE directly regulated the activity of ERK. Although LOE exerted inhibitory effect on melanin synthesis, we could not observe this effect in PD98059-treated α-MSH induced B16F10. These results strongly indicate that LOE is related to ERK activation and MITF degradation in anti-skin pigmentation. Hence, LOE should be utilized as a whitening agent of skin in the near future.

• Journal of Photoscience
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• v.9 no.2
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• pp.201-204
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• 2002

Epidermal cells produce a panel of antioxidants as well as cytokines after UVB irradiation, which counteract reactive oxygen species, however, how these antioxidants might regulate melanogenesis is unclear. An important constituent of the cellular antioxidant buffering system which controls the redox state of proteins is thioredoxin (TRX), a 13-kD protein that catalyzes thiol-disulfide exchange reactions, regulates activation of transcription factors, and possesses several other biological functions similar to cytokines. TRX suppressed the UVB-induced production and secretion of $\alpha$-melanocyte stimulating hormone ($\alpha$-MSH) and of adrenocorticotropic hormone (ACTH), and also suppressed proopiomelanocortin (POMC) mRNA expression by normal human keratinocyte (KC)s. Further, L-cysteine, N-acetyl-cysteine, $\alpha$-tocopheryl ferulate showed suppressive effect on UVB-induced POMC mRNA expression. However, TRX released from UVB-irradiated KCs stimulated melanogenesis by up-regulating MSH receptor expression and its binding activity in melanocyte (MC)s. UVB-induced KC derived cytokines such as IL1, IL6, and ET1 upregulated MSH-receptor binding ability as well as MCl-R mRNA expression in cultured normal human MCs. MCl-R has a tendency to be upregulated by UVB-induced KC-derived cytokines as well as by direct UVB irradiation. These results suggest that antioxidants such as TRX suppresses UVB induction of POMC, but in the case of MCl-R, this gene can be mainly in the trend of upregulation by UVB-induced KC-derived factors including TRX.

• Korean Journal of Pharmacognosy
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• v.33 no.4 s.131
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• pp.395-398
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• 2002

A chemical investigation of Peucedanum praeruptorum has resulted in the isolation of 3 khellactone derivatives, which have inhibitory effects on melanogenesis in Bl6 mouse melanoma cell lines. The khellactone derivatives were isolated from the crude extract of the roots of Pecedanum praeruptorum by a combination of adsorption chromatography and HPLC. The structlues of isolated compounds were identified as 3',4'- diangeloyl-cis-khellactone, 3'-angeloyl- 4'- senecioyl-cis-khel- lactone and,3', 4'-disenecioyl-cis-khellactone by $^1H\;NMR$, $^{13}C\;NMR$ and mass spectral studies and by comparisons of spectral data with reported literatures.

• YAKHAK HOEJI
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• v.49 no.5
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• pp.375-379
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• 2005

The green marine algae, Capsosiphon fulvescens is one of the important economic seaweeds cultured in Korea. In this study, we investigated the effects of Capsosiphon fulvescens on melanogenesis using B16 cells. Our results showed that Capsosiphon fulvescens significantly inhibits melanin synthesis and it reduces the activity of tyrosinase, the rate-limiting melanogenic enzyme. Western Blot analysis using anti-tyrosinase antibody revealed that Capsosiphon fulvescens ($10\~40\;{\mu}g/ml$ decreased tyrosinase protein levels. Cell proliferation was dose-dependently inhibited by 10, 20 and 40 ${\mu}g/ml$ Cap­sosiphon fulvescens, without cytotoxicity and morphological change. These results suggest that the depigmenting effect of Capsosiphon fulvescens is correlated with the suppression of tyrosinase activity and protein level, which are key enzymes for melanogenesis.

• Journal of Physiology & Pathology in Korean Medicine
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• v.19 no.4
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• pp.938-944
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• 2005

Down-regulation of melanin synthesis is required for recovery of pigmentary disorders and it is known that direct inhibitors of tyrosinase suppress melanin synthesis. We screened several oriental medicinal plants using B16/Fl0 cells and found that the methanolic extract of Cnidii Rhizoma down-regulated melanin synthesis effectively. Although the proliferation of Bl6/Fl0 cells was decresed by the methanolic extract of Cnidii Rhizoma, it did not appear necrosis. Bl6/F10 cells incubated with the methanolic extract of Cnidii Rhizoma showed reduced pigmentation and tyrosinase activity. Western blotting revealed that the amount of tyrosinase was decreased by the methanolic extract of Cnidii Rhizoma. These results suggest that the inhibitary effect of the methanolic extract of Cnidii Rhizoma on melanogenesis is due to the suppression of tyrosinase in Bl6/F10 cells and Cnidii Rhizoma is a candidate for an efficient whitening agent.

• Journal of Physiology & Pathology in Korean Medicine
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• v.24 no.2
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• pp.236-241
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• 2010

The purpose of this study was to investigate the mechanism of ethanol extract of Agrimonia pilosa Ledeb. (EAP)-reduced melanogenesis and diphenyl-picryl-hydrazyl (DPPH) radical scavenging activity. Agrimonia pilosa Ledeb., a perennial herbaceous plant, has been used as an antihemorrhagic, anthelminntic, and antiinflammatory agents in Chinese herbal medicine. In the present study, we observed that melanin synthesis and tyrosinase activity of B16F10 cells were significantly decreased by EAP. However, EAP could not suppress tyrosinase activity in the cell-free system, whereas kojic acid directly inhibited tyrosinase activity. Furthermore, EAP decreased the protein expression of tyrosinase, tyrosinase-related prootein 1 (TRP-1), and tyrosinase-related prootein 2 (TRP-2). EAP scavenged DPPH radical up to 41% with 100 ${\mu}g/m{\ell}$ concentration. These results suggest that the hypopigmentary effect of EPA was due to regulation of tyrosinase protein.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2022.09a
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• pp.110-110
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• 2022

Scrophularia koraiensis Nakai (S. koraiensis) has used its roots as traditional herbal medicine. Some research is reported to be effective in allergic inflammation and osteoporosis. In a present study, we conducted to investigate the bioactivity of the ethanol extract of S. koraiensis (ESK) on the inhibition of melanogenesis and the apoptosis of melanocytes. We analyzed Harpagoside of ESK by using LCMS and HPLC-PDA and investigated the regulation of ESK on reactive oxygen species. Also, the expressions of melanin synthesis-related factors and apoptosis-related factors were confirmed. As a result, the quantification results of quercetin and rutin in ESK were 77.2 and 7.4 mg/g. IC₅₀ on DPPH and ABTS radical scavenging activity is 33.1 and 9.5 ug/mL. ESK attenuated not only the expression of tyrosinase, TYRP-1, TYRP-2, and MITF in melanogenesis. It is thought that ESK may be effective in the inhibition of melanogenesis through MAPK cell signaling pathway in melanocytes. These study results suggest that ESK has the ability to inhibit melanin production and induce apoptosis.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2022.09a
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• pp.104-104
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• 2022

Dendropanax morbiferus H. Lev has been reported to have some pharmacologic activities and also interested in functional cosmetics. We found that the water extract of D. morbiferus leaves significantly inhibited tyrosinase activity and melanin formation in α-melanocyte stimulating hormone (MSH)-induced B16-F10 cells. D. morbiferus reduced melanogenesis-related protein levels, such as microphthalmia? associated transcription factor (MITF), TRP-1, and TRP-2, without any cytotoxicity. Two active ingredients of D. morbiferus, (10E)-9,16-dihydroxyoctadeca-10,17-dien-12,14-diynoate (DMW-1) and (10E)-(?)-10,17-octadecadiene-12,14-diyne-1,9,16-triol (DMW-2) were identified by testing the anti-melanogenic effects and then by liquid chromatography-tandem mass spectrometry (LC/MS/MS) analysis. DMW-1 and DMW-2 significantly inhibited melanogenesis by the suppression of protein kinase A (PKA)/cyclic AMP (cAMP)-responsive binding protein (CREB) and p38 MAPK phosphorylation. DMW-1 showed a better inhibitory effect than DMW-2 in α-MSH-induced B16-F10 cells. D. morbiferus and its active component DMW-1 inhibited melanogenesis through the downregulation of cAMP, p-PKA/CREB, p-p38, MITF, TRP-1, TRP-2, and tyrosinase. These results indicate that D. morbiferus and DMW-1 may be useful ingredients for cosmetics and therapeutic agents for skin hyperpigmentation disorders.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.30 no.1
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• pp.33-38
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• 2004

Melanin pigmentation in human skin is a major defense mechanism against ultraviolet light of the sun, but abnormal pigmentation such as freckles, liver spot could be a serious aesthetic problem. Nearly all studies are mainly concentrated on searching for the materials that have inhibitory activities on tyrosinase. In this work, to isolate phenolic compounds from Gastrodia elata, we purified the extract through solvent fractionation, column chromatography, and recrystallization. They were identified as 4-hydroxybenzyl alcohol 1, bis(4-hydroxyphenyl)methane 2, gastrodin (4-${\beta}$-D-glucopyranosyloxybenzyl alcohol) 3 on the base of spectroscopic evidences. In order to investigate their depigmentation effect, inhibitory activity against mushroom tyrosinase and inhibitory activity of melanin synthesis in B16 melanoma cells were evaluated in vitro. We have found that 4-hydroxybenzyl alcohol 1 and gastrodin (4- ${\beta}$-D-glucopyranosyloxybenzyl alcohol) 3 have no tyrosinase inhibitory activity, but inhibit the melanin synthesis in B16 melanoma cells. Tyrosinase inhibitory activities of bis(4-hydroxyphenyl)methane 2 (IC$\_$50/ = 400 $\mu\textrm{g}$/mL) and butanol fraction (IC$\_$50/ = 46 $\mu\textrm{g}$/mL) were lower/higher than that of arbutin (IC$\_$50/ = 114 $\mu\textrm{g}$/mL), but inhibitory activities of melanin synthesis in B16 melanoma cells were much higher than that of arbutin. Especially, tyrosinase inhibitory activities of isolated phenolic fraction (IC$\_$50/ = 2.37 $\mu\textrm{g}$/mL) from butanol fraction was very higher than that of arbutin (IC$\_$50/ = 114 $\mu\textrm{g}$/mL). Therefore, these results suggest that isolated phenolic compounds from Gastrodia elata have inhibitory activity against mushroom tyrosinase and inhibitory activity of melanin synthesis in 816 melanoma cells in vitro.

• Journal of Life Science
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• v.17 no.6 s.86
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• pp.873-875
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• 2007

Effects of hot-water extract from Sasa quelpaertensis leaf (HWES) on melanogenesis were investigated in B16/F10 mouse melanoma cells. HWES inhibited cellular tyrosinase activity and melanin biosynthesis in a dose-dependent manner. Western blotting analysis showed that HWES dose-dependently inhibited tyrosinase and tyrosinase related protein-1 expression. Also, HWES suppressed sustained ERK activation in a concentration-dependent manner, suggesting that HWES inhibits the melanin biosynthesis through the suppressive effect against pathway involving sustained ERK activation.