• Korean Journal of Food Science and Technology
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• v.48 no.1
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• pp.59-65
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• 2016

This study was carried out to investigate the total polyphenols, antioxidant activities, and melanin synthesis inhibition of several pear cultivars (Pyruspyrifolia). The total polypenolic content of five pear cultivars was high in the unripe developmental stage. Total polyphenolic content of the Chuwhangbae cultivar extract was higher than that of other pear cultivars regardless of the developmental stage. However, the total flavonoid content did not differ between cultivars or developmental stages. The phenolic compound, arbutin has an inhibitory effect on melanogenesis. Arbutin levels in pear cultivars declined as the fruit matured. The free radical scavenging activity of the extract also decreased as the fruit ripened. In B16F10 mouse melanoma cells, most of the cultivar extracts inhibited melanin synthesis by about 50% at a $100{\mu}g{\cdot}mL^{-1}$ concentration, except in the Gamcheonbae extract until 90 days after full bloom. We have confirmed that the extract of pear cultivars have antioxidant activity and skin-whitening effects.

• IMMUNE NETWORK
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• v.3 no.1
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• pp.53-60
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• 2003

Background: The role of macrophages in tumor angiogenesis is known to be the production of angiogenic cytokines and growth factors including TNF-${\alpha}$. Recently, macrophage also can produce the INF-${\gamma}$ that is being studied to be involved in angiogenic inhibition. Thus, the importance of macrophages in tumor angiogenesis is might being an angiogenic switch. Thus, the hypothesis tested here is that TNF-${\alpha}$ can modulate the INF-${\gamma}$ production in the macrophages from tumor environment as a part of tumor angiogenic switch. Methods: Macrophages in tumor environment were obtained from the peritoneal cavity of C57BL/6 mice injected with B16F10 melanoma cell line for 6 or 11 days. $Mac1^+$-macrophages were purified using magnetic bead ($MACs^{TM}$; Milteny Biotech, Germany) and cultured with various concentrations of TNF-${\alpha}$ for various time points at $37^{\circ}C$. The supernatants were analyzed for IFN-${\gamma}$ or VEGF by ELISA kit (Endogen, Woburn, MA). Results: Residential macrophages from the peritoneal cavity did not respond to LPS or TNF-${\alpha}$ to produce INF-${\gamma}$. However, the cells from tumor environment produced IFN-${\gamma}$ as well as VEGF and upregulated by the addition of LPS or TNF-${\alpha}$. RT-PCR analysis revealed the external TNF-${\alpha}$-induced IFN-${\gamma}$ gene expression in the macrophages from tumor environment. Conclusion: The overall data suggest that the macrophages in tumor environment might have an important role not only in angiogenic signal but also in anti-angiogenic signal by producing related cytokines. And TNF-${\alpha}$ might be a key cytokine in tumor angiogenic switch.

• Journal of Physiology & Pathology in Korean Medicine
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• v.21 no.5
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• pp.1243-1249
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• 2007

The aim of this study was to investigate the effect of ethanol extract of Fagopyrum esculentum on the melanogenesis. To determine whether ethanol extract of Fagopyrum esculentum suppress melanin synthesis in cellular level, B16F10 melanoma cells were cultured in the presence of different concentrations of Fagopyrum esculentum ethanol extract. In the present study, we examined the effects of Fagopyrum esculentum ethanol extract on cell proliferation, melanin contents, tyrosinase activity, expression of melanogenic enzyme proteins including tyrosinase, tyrosinase-related protein 1 (TRP-1) and tyrosinase-related protein 2 (TRP-2). Cell proliferation was slightly increased by treatment with ethanol extract of Fagopyrum esculentum $(25-200 {\mu}g/m{\ell}).$ The ethanol extract of Fagopyrum esculentum effectively suppressed melanin contents at a dose of $100 {\mu}g/m{\ell}).$ It was observed that the color of cell pellets was totally whitened compared with the control. The ethanol extract of Fagopyrum esculentum inhibited tyrosinase activity, regulate melanin biosynthesis as the key enzyme in melanogenesis. Using western blot analysis, the ethanol extract of Fagopyrum esculentum dose-dependently decreased tyrosinase and TRP-1 protein levels, and tyrosinase and TRP-1 were detected in similar manner. ${\alpha}-MSH$ leads to a stimulation of melanin synthesis through increase of tyrosinase activity, melanin contents and cytoplasmic dendricity. In this study, ethanol extract of Fagopyrum esculentum down-regulated the ${\alpha}-MSH$-induced tyrosinase activity, melanin contents and cytoplasmic dendricity. Regarding protein levels of the melanogenic enzymes, the amounts of tyrosinase and TRP-1 was increased after incubation with a-MSH. The treatment of ethanol extract of Fagopyrum esculentum decreased the ${\alpha}-MSH$-induced expression levels of tyrosinase and TRP-1. These results suggest that the ethanol extract of Fagopyrum esculentum exerts its depigmenting effects through the suppression of tyrosinase, TRP-1 and cytoplasmic dendricity. And it may be a potent depigmetation agent in hyperpigmentation condition.

• Proceedings of the Korean Society of Applied Pharmacology
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• 2003.11a
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• pp.98-98
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• 2003

${\beta}$-(1,3)-D-Glucans have been known to exhibit antitumor and antimicrobial activities. The presence of dectin-1,${\alpha}$, ${\beta}$-glucan receptor of dendritic cell, on macrophage has been controvertial. RT-PCR analysis led to the detection of dectin-1${\alpha}$ and ${\beta}$ in murine macrophage Raw264.7 cell line. Among the various organs of mouse, dectin-1${\alpha}$ and ${\beta}$ were detected in the thymus, lung, spleen, stomach and intestine. To analyze gene expression modulated by ${\beta}$-glucan treated murine Raw264.7 macrophage, total mRNA was applied to cDNA microarray to interrogate the expression of 7,000 known genes. cDNA chip analysis showed that ${\beta}$-glucan of P. osteatus increased gene expressions of immunomodulating genes, membrane antigenic proteins, chemokine ligands, complements, cytokines, various kinases, lectin associated genes and oncogenes in Raw 264.7 cell line. When treated with ${\beta}$-glucan of P. osteatus and LPS, induction of gene expression of TNF-${\alpha}$ and IFN-R1 was confirmed by RT-PCR analysis. Induction of TNF-R type II expression was confirmed by FACS analysis. IL-6 expression was abolished by EDTA in ${\beta}$-glucan and LPS treated Raw264.7 cell line, indicating that ${\beta}$-glucan binds to dectin-l in a Ca$\^$＋＋/ -dependent manner. To increase antitumor efficacy of ${\beta}$-glucan, ginsenoside Rh2 (GRh2) was co-treated with ${\beta}$-glucan in vivo and in vitro tests. IC$\sub$50/ values of GRh2 were 20 and 25 $\mu\textrm{g}$/$m\ell$ in SNU-1 and B16 melanoma F10 cell line, respectively. Co-treatment with ${\beta}$-glucan and GRh2 showed synergistic antitumor activity with cisplatin and mitomycin C both in vitro and in vivo. Single or co-treatment with ${\beta}$-glucan and GRh2 increased tumor bearing mouse life span. Co-treatment with ${\beta}$-glucan and GRh2 showed more increased life span with mitomycin C than that with cisplatin. Antitumor activities were 67% and 72 % by co-injection with ${\beta}$-glucan and GRh2 in the absence or presence of mitomycin C, respectively.

• Journal of the Korean Society of Food Science and Nutrition
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• v.42 no.4
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• pp.550-555
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• 2013

Extrinsic skin aging is characterized by the loss of skin tone and resilience, irregular pigmentation, and deep wrinkles. The aim of this study was to investigate the effects of Ehwa Makgeolli containing oriental herbs (Glycyrrhiza uralensis Fisch., Lycium chinense MILL., Morus alba L., and Saururus chinensis Baill) on skin whitening and wrinkling in human skin cells. We prepared Makgeolli extracts (HEE) with 70% ethanol. HEE significantly inhibited in vitro mushroom tyrosinase activity and reduced the cellular and secreted melanin content of mouse melanoma melanocytes (B16F1 cells). HEE down-regulated the protein expression of tyrosinase related protein (TRP)-1/-2, a key player in melanogenesis. Treatment with HEE in human keratinoctyes (HaCaT cells) inhibited the proteolytic activities of matrix metalloproteinase (MMP)-2/-9 in a dose-dependent manner and dramatically reduced the expression of MMP-2/-9. In addition, HEE attenuated lipopolysaccharide (LPS)-induced nitric oxide production in murine macrophages (RAW264.7 cells). These results indicate that HEE may be a great cosmeceutical ingredient for its whitening, anti-wrinkle, and anti-inflammatory effects.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.38 no.4
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• pp.277-287
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• 2012

To develop new therapeutic materials to prevent hair loss and enhance hair growth, we developed a medicinal herbal complex extract (MHCE) using 23 herbs traditionally used in oriental medicine. Medicinal Herbal complex extract was consist of Angelica gigas Nakai, Psoralea corylifolia Linne, Biota orientalis Endlicher, and Eclipta prostrata Linne, Rehmannia glutinosa Liboschitz var. purpurea Makino, Ligustrum lucidum Aiton, Polygonum multiflorum Thunberg, and Sesamum indicum Linne, Sophora angustifolia Sieboldet Zuccarini, Angelica dahurica Benthamet Hooker, and Leonurus sibiricus Linne, Salvia miltiorrhiza Bunge, Prunus persica Batsch, Commiphora molmol Engler, Chrysanthemum indicum Linne, Boswellia carterii Birdwood, Panax ginseng C. A. Meyer, Cnidium officinale Makino, Albizia julibrissin Durazzini, and Corydalis ternata Nakai that have traditionally been used for treating hair loss, preventing gray hair, anti-inflammation, and blood circulation in oriental medicine. In addition, we examined the hair growth effect of MHCE in vitro and in vivo. In vitro, we evaluated the effects of MHCE on cultured HFDPC, HaCaT cells, and murine embryonal fibroblasts (NIH3T3 cells). Also, we evaluated the ability of MHCE to prevent gray hair on murine melanoma cells (B16F1 cells). The hair growth-promoting effect of MHCE in vitro was also observed in vivo using C57BL/6 mice. Our results showed that MHCE significantly increased the proliferation of HFDPC (175 % proliferation at $50{\mu}g/mL$), HaCaT cells (133 % proliferation at $20{\mu}g/mL$), and NIH3T3 cells (120 % proliferation at $50{\mu}g/mL$). MHCE also showed consistent melanogenesis in B16F1 cells (154 % melanin synthesis at $50{\mu}g/mL$). Moreover, MHCE showed potential for hair growth stimulation in C57BL/6 mice experiments (98 % hair growth area on 4 weeks). These results indicate that MHCE may be a good candidate for promotion of hair growth.

• Korean Journal of Plant Resources
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• v.34 no.1
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• pp.37-43
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• 2021

Angelica gigas Nakai (AGN) is a perennial plant belonging to the family Apiaceae. Its root has been utilized as a traditional medicine especially in Korea. This study was carried out to evaluate the potential use of extracts from AGN root parts as a cosmetic material. The dried AGN roots are divided into body (B), thick root (TkR), medium root(MR) and thin root (TnR) according to their diameter before cutting into medicine. B, TkR and MR of AGN are combined and used as medicinal herbs (MH). The extracts from AGN each root part (B, TkR, MR, TnR, MH) were used to test the effect on cell viability using MTS assay and to examine inhibitory effect on melanin accumulation in B16F10 melanoma cells. All extracts (50 - 200 ㎍/mL) from the each root part did not affect the cell viability. And inhibitory effect of all root extracts (200 ㎍/mL) on melanin accumulation was 12-19%. Especially, TnR showed similar inhibitory effect on melanin accumulation to MH. In addition, DPPH and ABTS free radical scavenging activity were higher in the TnR extract compared to MH. This study showed that the TnR extract exhibit high inhibitory effect on melanin accumulation and antioxidant activity compared to MH, suggesting that TnR extract has potential as a cosmetic ingredient.

• 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.