• Journal of the Society of Cosmetic Scientists of Korea
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• v.45 no.1
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• pp.49-56
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• 2019

Pueraria thomsonii Benth. as a medicinal ingredient, has been traditionally used in Chinese medicine to treat fever, acute dysentery, diarrhea, diabetes, and cardiovascular disease. The effects of P. thomsonii flower on skin have not been reported yet. In this study, the whitening effect of P. thomsonii flower was verified using B16F1 melanoma cells and HS68 fibroblasts. P. thomsonii flower extract reduced melanin contents of B16F1 cells in a dose-dependent manner. To identify its active components, we analyzed P. thomsonii flower extract using high performance liquid chromatography (HPLC). As a result, we identified three major isoflavones of tectorigenin, tectoridin, and tectorigenin 7-O-xylosylglucoside. At a non-cytotoxic concentration, the three components also reduced melanin contents of B16F1 cells in a dose-dependent manner. The depigmentation effects were attributed to the reduced gene expression of tyrosinase and microphthalmia-associated transcription factor (MITF). In order to elucidate another depigmentation mechanism, their effects on DKK-1, a fibroblast-derived depigmentation factor, was determined in HS68 cells. As a result, P. thomsonii flower extracts, tectoridin and tectorigenin 7-O-xylosylglucoside, reduced DKK-1 gene expression, while tectorigenin increased DKK-1 gene expression in a dose-dependent manner. These results suggest that tectorigenin can be used as an effective whitening agent that inhibit melanin synthesis in melanocytes and promote the secretion of depigmentation factor from fibroblasts.

• Journal of Life Science
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• v.32 no.5
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• pp.355-361
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• 2022

Melanin pigments are the main cause of skin color. They are produced in melanocytes and then transferred to keratinocytes, which eventually gives the skin surface a variety of colors. Although many skin-lightening or depigmenting agents have been developed, the demand for materials to reduce pig- mentation is still increasing. Here, we tried to find materials for skin-lightening or depigmentation using natural compounds and found that mistletoe (Viscum album var. coloratum) extracts (ME) had an inhibitory effect on tyrosinase activity. As a result, ME significantly reduced pigmentation in human primary melanocytes. In addition, a promoter reporter assay revealed that ME inhibited the transcription of microphthalmia-associated transcription factor (MITF), melanophilin (MLPH), tyrosinase-related protein-2 (TRP-2), and tyrosinase (TYR) genes in HM3KO melanoma cells. In addition, ME decreased the protein level for pigmentation-related molecules, such as TYR and TRP-1. Furthermore, it markedly inhibited the melanogenesis of zebrafish embryos, an in vivo evaluation model for pigmentation. To elucidate the action mechanism of ME, we investigated its effects on intracellular signaling. Eventually, the ME dramatically decreased the phosphorylation of the cAMP responsive element binding protein (CREB), AKT, and ERK. The data suggest that ME may inhibit the melanogenesis pathway by regulating the signaling pathway related to pigmentation. Taken together, these data propose that ME can be developed as a depigmenting or skin-lightening agent.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.36 no.3
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• pp.175-182
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• 2010

In this study, the antioxidative effects, inhibitory effects on tyrosinase of Cedrela sinensis extracts were investigated. The ethyl acetate fraction of extract ($3.54\;{\mu}g/mL$) and aglycone fraction of extract ($2.15\;{\mu}g/mL$) showed more excellent free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activity ($FSC_{50}$) than the activity of (+)-$\alpha$-tocopherol ($8.98\;{\mu}g/mL$). Reactive oxygen species (ROS) scavenging activities ($OSC_{50}$) of Cedrela sinensis extracts on ROS generated in $Fe^{3+}$-EDTA/$H_2O_2$ system were investigated using the luminol-dependent chemiluminescence assay. The ethyl acetate fraction of extract ($0.15\;{\mu}g/mL$) and aglycone fraction of extract ($0.12\;{\mu}g/mL$) showed 10 times more excellent ROS scavenging activity than activity of L-ascorbic acid ($1.50\;{\mu}g/mL$). The protective effects of fractions of Cedrela sinensis on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The ethyl acetate fraction of extract and aglycone fraction of extracts suppressed photohemolysis in a concentration dependent manner ($5{\sim}25\;{\mu}g/mL$). The inhibitory effect of Cedrela sinensis extracts on tyrosinase was investigated to assess their whitening efficacy. Inhibitory effects ($IC_{50}$) on tyrosinase were determined with ethyl acetate fraction of Cedrela sinensis extract ($48.00\;{\mu}g/mL$) and aglycone fraction of extract ($5.88\;{\mu}g/mL$). The aglycone fraction showed 40 times more remarkable tyrosinase inhibitory effect than whitening agent, arbutin ($226.88\;{\mu}g/mL$) These results indicate that fractions of Cedrela sinensis can be used as antioxidants in biological systems, particularly skin exposed to UV radiation by scavenging $^1O_2$ and other ROS, and protect cellular membranes against ROS. The fractions of Cedrela sinensis can be applicable to new functional cosmetics for antioxidant and whitening.

• Proceedings of the Korean Society of Applied Pharmacology
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• 2007.11a
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• pp.79-92
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• 2007

Oxidative stress have known to be a risk factor for the degenerative processes and closely related to a lot of diseases. It is well established that antioxidants are good in protection and therapeutic means against oxidative damage. There is increasing interest in natural antioxidants and many natural antioxidants have been found and utilized as the possible protection for various diseases and skin aging. We have screened natural antioxidant agents for cosmeceuticals, nutraceuticals, and drugs as therapeutic and preventive means against oxidative stress, and have developed a number of novel antioxidants from various natural sources. A novel melanin synthesis inhibitor, Melanocin A, isolated from the metabolite of a fungal strain Eupenicillium shearii F80695 inhibited mushroom tyrosinase and melanin biosynthesis of B16 melanoma cells with $IC_{50}$ value of 9.0 nM and MIC value of $0.9\;{\mu}M$, respectively. Melanocin A also exhibited potent antioxidant activity by scavenging of DPPH and superoxide anion radicals. UV was found to increase the level of hydrogen peroxides and other reactive oxygen species (ROS) in skin tissues. This increase in ROS may not only alter the structure and function of many genes and proteins directly but may also modulate their expressions through signal transduction pathways and, ultimately, lead to skin damage. We investigated the effect of Melanocin A on UV-induced premature skin aging. Firstly, the effect of Melanocin A on UV-induced matrix metalloproteinase (MMP)-9 expression in an immortalized human keratinocyte cell line, HaCaT in vitro was investigated. Acute UV irradiation induced MMP-9 expression at both the mRNA and protein levels and Melanocin A suppressed this expression in a dose-dependent manner. We then investigated UV-induced skin changes in hairless mice in vivo by Melanocin A. Chronic exposure of hairless mouse dorsal skin to UV increased skin thickness and induced wrinkle formation and the gelatinase activities of MMP-2 and MMP-9. Moreover, Melanocin A significantly suppressed UV-induced morphologic skin changes and MMP-2 and MMP-9 expression. These results show that Melanocin A can prevent the harmful effects of UV that lead to skin aging. Therefore, we suggest that Melanocin A should be viewed as a potential therapeutic agent for preventing and/or treating premature skin aging. Terrein is a bioactive fungal metabolite isolated from Penicillium species. Terrein has a relatively simple structure and can be easily synthesized. However, the biologic effects of terrein are comparatively unknown. We found for the first time that terrein potently inhibit melanin production in melanocytes and has a strong hypopigmentary effect in a spontaneously immortalized mouse melanocyte cell line, Mel-Ab. Treatment of Mel-Ab cells with terrein (10-100 mM) for 4 days significantly reduced melanin levels in a dose-dependent manner. In addition, terrein at the same concentration also reduced tyrosinase activity. We then investigated whether terrein influences the extracellular signal-regulated protein kinase (ERK) pathway and the expression of microphthalmia-associated transcription factor (MITF), which is required for tyrosinase expression. Terrein was found to induce sustained ERK activation and MITF down-regulation, and luciferase assays showed that terrein inhibits MITF promoter activity in a dose-dependent manner. To elucidate the correlation between ERK pathway activation and a decreased MITF transcriptional level, PD98059, a specific inhibitor of the ERK pathway, was applied before terrain treatment and found to abrogate the terrein-induced MITF attenuation. Terrein also reduced the tyrosinase protein level for at least 72 h. These results suggest that terrain reduces melanin synthesis by reducing tyrosinase production via ERK activation, and that this is followed by MITF down-regulation.

• Journal of the Korean Applied Science and Technology
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• v.28 no.4
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• pp.482-490
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• 2011

In this study, the antioxidative effects and inhibitory effects on tyrosinase and elastase of Sorbus commixta (S. commixta) twig extracts were investigated. The aglycone fraction of S. commixta twig extract showed the prominent free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activity($FSC_{50}$, $13{\mu}g/mL$). Reactive oxygen species (ROS) scavenging activities of S. commixta twig extracts on ROS generated in $Fe^{3+}-EDTA/H_2O_2$ system were investigated by the luminol-dependent chemiluminescence assay. The 50 % ethanol extract among extracts showed the most prominent ROS scavenging activity ($OSC_{50}$, $0.189{\mu}g/mL$). The cellular protective effects of extract/fractions of S. commixta twig on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The 50 % ethanol extract and ethyl acetate fraction showed the cellular protective effects against ROS in a concentration dependent manner ($5{\sim}50{\mu}g/mL$). The inhibitory effect of S. commixta twig extract on tyrosinase was investigated to assess the whitening efficacy. The ethyl acetate ($IC_{50}$, $113.2{\mu}g/mL$) and aglycone fraction($IC_{50}$, $105.3{\mu}g/mL$) on tyrosinase showed more remarkable inhibitory effect than arbutin($IC_{50}$, $226.88{\mu}g/mL$), known as the whitening agent. The inhibitory effect of aglycone fraction ($IC_{50}$, $6.9{\mu}g/mL$) on elastase was simillar to quercetin($IC_{50}$, $6.1{\mu}g/mL$), flavonoid known as reference compound. These results indicate that extract/fractions of S. commixta twig can function as antioxidants in biological systems, particularly skin exposed to UV radiation by scavenging $^1O_2$ and other ROS, and protect cellular membranes against ROS. S. commixta twig extracts can be applicable to new functional cosmetics for anti-aging products.

• Korean Journal of Medicinal Crop Science
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• v.21 no.5
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• pp.361-371
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• 2013

This study was performed to determine the antimelanogenic effect and tyrosinase inhibitory activities of anthocyanin rich fraction (AN-SLP) from Liriope platyphylla Wang et Tang seeds. Anthocyanins isolated from L. platyphylla seeds revealed the presence of four major anthocyanin components, which were tentatively identified as delphinidin-3-Oglucoside, delphinidin-3-O-rutinoside, petunidin-3-O-rutinoside, and malvidin-3-O-rutinoside using semipreparative HPLC, $^1H$-NMR, $^{13}C$ NMR, FAB-MS and LC/ES-MS. The inhibitory effect of AN-SLP on tyrosinase activity was studied using in vitro (against mushroom tyrosinase) and ex vivo (against B16 melanoma cell tyrosinase) models. Cellular tyrosinase activity was decreased by AN-SLP treatment in B 16 melanoma cells through dose dependent manner, but AN-SLP did not inhibit mushroom tyrosinase and L-DOPA oxidation directly. AN-SLP showed melanin inhibition by 53.2% at 50 ${\mu}g/m{\ell}$ which was 0.7 times more efficient than the antimelanogenic effect of commercial arbutin and kojic acid (36.5%) also did not show cell toxicity. Additionally, AN-SLP inhibited the activity of ${\alpha}$-glucosidase and the glycosylation of tyrosinase in melanoma cell. The resulting unsaturated glycosylation of tyrosinase makes it unstable and disturb correct transportation. From theses results, we conclude that AN-SLP could be used as anti-melanogenic agent for skin whitening.

• Korean Journal of Food Science and Technology
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• v.54 no.3
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• pp.280-287
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• 2022

The loquat (Eriobotrya japonica) is a fruit tree belonging to the Rosaceae family. Loquat fruit extracts from three cultivars (Tanaka, Mogi, and Jinwang) were prepared using absolute methanol and homogenization and ultrasound sonication procedures. We measured the total phenolic and flavonoid content of the fruit extracts and their antioxidant capacities. In addition, we evaluated tyrosinase activity and anti-melanogenic effects in B16F1 melanoma cells. The fruit extract from cv. Tanaka had the highest total phenolic content and showed the highest antioxidant capacity in the ABTS assay. The loquat fruit extract from cv. Jinwang had the highest total flavonoid content and showed the highest antioxidant capacity in the DPPH assay. Loquat fruit extracts from the Tanaka and Jinwang cultivars effectively inhibited mushroom tyrosinase activity. The loquat fruit extracts reduced intracellular oxidative stress in B16F1 melanoma cells. Treating B16F1 melanoma cells with loquat fruit extract from cv. Tanaka at a concentration of 125 ㎍/mL effectively inhibited melanin synthesis. However, treating the B16F1 melanoma cells with loquat fruit extracts from the Jinwang and Mogi cultivars did not. These results suggest that loquat fruit extracts from the Tanaka cultivar may serve as potential sources of antioxidants and act as a skin-whitening agent.

• Toxicological Research
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• v.33 no.1
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• pp.55-62
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• 2017

We evaluated the antioxidant activity and anti-melanogenic effects of Oenothera laciniata methanol extract (OLME) in vitro by using melan-a cells. The total polyphenol and flavonoid content of OLME was 66.3 and 19.0 mg/g, respectively. The electron-donating ability, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical-scavenging activity, and superoxide dismutase (SOD)-like activity of OLME ($500{\mu}g/mL$) were 94.5%, 95.6%, and 63.6%, respectively. OLME and arbutin treatment at $50{\mu}g/mL$ significantly decreased melanin content by 35.5% and 14.2%, respectively, compared to control (p < 0.05). OLME and arbutin treatment at $50{\mu}g/mL$ significantly inhibited intra-cellular tyrosinase activity by 22.6% and 12.6%, respectively, compared to control (p < 0.05). OLME ($50{\mu}g/mL$) significantly decreased tyrosinase, tyrosinase-related protein-1 (TRP-1), TRP-2, and microphthalmia-associated transcription factor-M (MITF-M) mRNA expression by 57.1%, 67.3%, 99.0%, and 77.0%, respectively, compared to control (p < 0.05). Arbutin ($50{\mu}g/mL$) significantly decreased tyrosinase, TRP-1, and TRP-2 mRNA expression by 24.2%, 42.9%, and 48.5%, respectively, compared to control (p < 0.05). However, arbutin ($50{\mu}g/mL$) did not affect MITF-M mRNA expression. Taken together, OLME showed a good antioxidant activity and anti-melanogenic effect in melan-a cells that was superior to that of arbutin, a well-known skin-whitening agent. The potential mechanism underlying the anti-melanogenic effect of OLME was inhibition of tyrosinase activity and down-regulation of tyrosinase, TRP-1, TRP-2, and MITF-M mRNA expression.

• Journal of Microbiology and Biotechnology
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• v.25 no.12
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• pp.2011-2015
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• 2015

Ginsenoside Rb2 (Gin-Rb2) was purified from the fruit extract of Panax ginseng. Its chemical structure was measured by spectroscopic analysis, including HR-FAB-MS, ¹H-NMR, and IR spectroscopy. Gin-Rb2 decreased potent melanogenesis in melan-a cells, with 23.4% at 80 μM without cytotoxicity. Gin-Rb2 also decreased tyrosinase and MITF protein expression in melan-a cells. Furthermore, Gin-Rb2 presented inhibition of the body pigmentation in the zebrafish in vivo system and reduced melanin contents and tyrosinase activity. These results show that Gin-Rb2 isolated from P. ginseng may be an effective skin-whitening agent via the in vitro and in vivo systems.

• Journal of Physiology & Pathology in Korean Medicine
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• v.24 no.4
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• pp.674-680
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• 2010

The purpose of this study was to investigate the effects of Fructus Ligustri Lucidi (FLL) on the process of melanogenesis. Cell viability of B16F10 cells was measured by MTT assay and melanin content was assessed using the method of Hosoi with some modifications. Methanol extract of Fructus Ligustri Lucidi (MFLL) significantly inhibited melanin synthesis in a concentration-dependent manner, and it reduced the activity of tyrosinase, the rate-limiting melanogenic enzyme. Additionally, $\alpha$-melanocyte stimulation hormone ($\alpha$-MSH)-induced hyperpigmentation was down-regulated by MFLL. MFLL was fractionated by organic solvent. In the present study, Hexane extract of Fructus Ligustri Lucidi (HFLL) inhibited tyrosinase activity and melanin production of B16F10 cells in the absence or presence of $\alpha$-MSH. Ethyl acetate, butanol and water layers did not affect tyrosinase activity and melanin production. Meanwhile ethyl acetate, butanol and water layers showed DPPH free radical scavenging activity. These results suggest that extract of FLL could be used as functional biomaterial in developing a skin whitening agent having the antioxidant activity.