• The Korean Journal of Physiology and Pharmacology
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• v.21 no.3
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• pp.287-292
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• 2017

Vitiligo is an intriguing depigmentary disorder and is notoriously difficult to be treated. The ultimate goal of vitiligo treatment is to replenish the lost melanocytes by immigration from hair follicle and to restore the normal function of melanogenesis by residual melanocytes. There are two types of topical calcineurin inhibitors called tacrolimus and pimecrolimus, and are recommended as the first-line treatments in vitiligo. Although pimecrolimus is efficacious for the repigmentation of vitiligo, its intrinsic mechanisms have never been investigated in vitro. This research aimed to study the ability of pimecrolimus on stimulating melanogenesis, melanocyte migration and MITF (microphthalmia associated transcription factor) protein expression. Results showed that pimecrolimus at the dosages of 1, 10, $10^2$nM were neither mitogenic nor cytotoxic to melanocytes. The addition of pimecrolimus at 10, $10^2$ and $10^3nM$ significantly increased intracellular tyrosinase activity, which was consistent with the elevated content of melanin content at the same concentrations. The peak effect was seen at 72 h in response to $10^2$nM pimecrolimus. Results of the wound scratch assay and Transwell assays indicate that pimecrolimus is effective in facilitating melanocyte migration on a collagen IV-coated surface. In addition, MITF protein yield reached the highest by pimecrolimus at $10^2nM$. In brief, pimecrolimus enhances melanin synthesis as well as promotes migration of melanocytes directly, possibly via their effects on MITF protein expression.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.26 no.1
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• pp.1-18
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• 2013

Objective: Recently the demands for the effective and safe depigmentative and anti-aging agents of the skin have increased due to the medical, pharmaceutical and cosmetic reasons. The purpose of this study is to investigate the MKG(Methanol Extract of Kaempferia galanga) and their dermal bioactivity properties related to cosmeceuticals such as depigmentation. Methods: We assessed inhibitory effects of MKG on melanin production in B16/F10 melanoma cells, on mushroom tyrosinase activity, effects of MKG on the expression tyrosinase, TRP-1, TRP-2, GSK-$3{\beta}$, CREB, MITF in B16/F10 melanoma cells without cytotoxicity range. Cell viability was measured by MTT assay and tyrosinase activity was assessed using by DOPA staining, western-blot analysis. We measured inhibition of melanin synthesis and tyrosinase activity by down-regulation of melanogenic enzyme expressions in ${\alpha}$-MSH induced melanogenesis B16/F10 melanoma cells. Results: MKG inhibited tyrosinase-activity, total melanin contents and dendrite out-growth. MKG inhibited melanogenesis by down-regulation of tyorsinase, TRP-1, TRP-2, CREB, and MITF in B16/F10 cells. The treatment with MKG at the 12.5, $25{\mu}g/ml$ level significantly inhibited the melanin synthesis induced ${\alpha}$-MSH in B16/F10 melanoma cells compared with untreated control. Conclusion: These results suggest that MKG inhibit melanin biosynthesis which is involved in hyper-pigmentation. So MKG is considered to be used as a whitening components reducing cytotoxicity.

• Korean Journal of Pharmacognosy
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• v.52 no.3
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• pp.143-148
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• 2021

This study was performed to elucidate the inhibitory effects of Alveopora japonica extract on melanin synthesis by measuring the levels of cell viability, mRNA expression, tyrosinase activity, and melanin production in the B16F10 cell line. The effects of A. japonica extract on tyrosinase-related protein 1 (TYRP1), TYRP2, tyrosinase (TYR), and microphthalmia-associated transcription factor (MITF) mRNA expression levels and melanin content were determined. Quantitative real-time RT-PCR show that A. japonica extract decrease the mRNA expression levels of TYRP1, TYRP2, TYR, and MITF in B16F10 cell line, resulting in lower levels of melanin production compared to α-MSH-treated B16F10 cells. Tyrosinase activity assays reveal that A. japonica extract decrease melanin production in B16F10 cells. These results demonstrate the whitening effects of A. japonica extract on B16F10 cells; thus, A. japonica extract is a potent ingredient for skin whitening. Further research is needed on the mechanism of action of A. japonica extract. Such research will benefit not only cosmetics, but also the health food and medical industries.

• The Korean Journal of Physiology and Pharmacology
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• v.18 no.4
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• pp.307-311
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• 2014

In this study, we isolated scopoletin from Cirsium setidens Nakai (Compositae) and tested its effects on melanogenesis. Scopoletin was not toxic to cells at concentrations less than $50{\mu}M$ and increased melanin synthesis in a dose-dependent manner. As melanin synthesis increased, scopoletin stimulated the total tyrosinase activity, the rate-limiting enzyme of melanogenesis. In a cell-free system, however, scopoletin did not increase tyrosinase activity, indicating that scopoletin is not a direct activator of tyrosinase. Furthermore, Western blot analysis showed that scopoletin stimulated the production of microphthalmia-associated transcription factor (MITF) and tyrosinase expression via cAMP response element-binding protein (CREB) phosphorylation in a dose-dependent manner. Based on these results, preclinical and clinical studies are needed to assess the use of scopoletin for the treatment of vitiligo.

• 한국생물공학회:학술대회논문집
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• 2005.10a
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• pp.191-191
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• 2005

• 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.

• Archives of Plastic Surgery
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• v.38 no.6
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• pp.725-732
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• 2011

Purpose: Skin grafting is one of the most commonly used methods in reconstructive plastic surgery field, but complications such as color change, contracture or hypertrophy are common problems. However, pathophysiology of the color change after skin graft is not yet determined and no animal model is established. Methods: Full thickness skin grafts were performed on the dorsum of C57BL/6 mice. Serial chronological gross inspection for color change and pigmentation were examined. Melanin pigments were traced by Fontana-Masson staining and semi-quantitative analysis was performed. In addition, immunohistochemical staining of S-100, Micropthalmia related Transcription Factor (MITF) and Melan-A antibodies were also performed to observe melanocytes and their changes. Results: After skin graft, color change and pigment spots were observed in the graft. Fontana-Masson staining showed melanin pigments in the epidermal and dermal layers in all mice. Immunohistochemistry staining to S-100, MITF, Melan-A antibodies showed melanocytes at the basal layer of epidermis and dermis. Conclusion: In conclusion, we have established an animal model for skin pigmentation after skin graft. We believe this study may be useful in understanding of the behavior of melanocytes after skin graft.

• KSBB Journal
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• v.32 no.3
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• pp.233-237
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• 2017

Melanin is one of the most important factors affecting skin color. Melanogenesis is the bioprocess of melanin production by melanocytes in the skin and hair follicles and is mediated by several enzymes, such as tyrosinase, tyrosinase related protein (TRP)-1, and TRP-2, MITF. In this study, we investigated the effect of Artemisia fukudo Makino extracts on tyrosinase activity and melanin production as natural products of whitening functional cosmetics. Melanin content in murine B16F10 melanoma cells were decreased by Artemisia fukudo Makino extracts in a dose-dependently. In addition, the inhibition of tyrosinase activity of Artemisia fukudo Makino extracts showed to decrease tyrosinase activity as the concentration of ${\alpha}-MSH$ was increased. Furthermore, western blot analysis revealed that Artemisia fukudo Makino extracts significantly downregulated the expression of tyrosinase, TRP-1 which treat of ${\alpha}-MSH-induced$ melanogenesis in murine B16F10 melanoma cells. As a result, Artemisia fukudo Makino extract showed functionalities as an effective whitening agent to inhibit melanin formation.

• Journal of Applied Biological Chemistry
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• v.64 no.4
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• pp.447-451
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• 2021

In this study, we discovered for the first time that linarin, a flavonoid compound, enhances melanin biosynthesis in B16F10 cells, and subsequently elucidated the underlying mechanism of linarin-induced melanogenesis. Linarin showed no cytotoxicity at a concentration of 42 μM and significantly increased intracellular tyrosinase activity and melanin content in B16F10 cells. Mechanistic analysis showed that linarin increased the expression of tyrosinase, tyrosinase-related protein 1 (TRP-1), and microphthalmia-associated transcription factor (MITF) that are related to melanogenesis. Moreover, linarin decreased the phosphorylation of extracellular signal-regulated kinase (ERK) and protein kinase B (AKT). Finally, we evaluated the effect of the structure-activity relationship of linarin and its aglycone on melanogenesis. The results indicated that linarin enhances the expression of melanogenic proteins by activating MITF expression via the modulation of mitogen-activated protein kinase (MAPK), phosphatidylinositol 3-kinase (PI3K), and protein kinase B signaling pathways in B16F10 cells, thereby enhancing melanogenesis.

• The Journal of Korean Obstetrics and Gynecology
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• v.29 no.2
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• pp.66-82
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• 2016

Objectives : This study was conducted to evaluate the effect of Kanghwalsokdan-tang Gamibang water extract (KSG) on osteoporosis. Methods : RANKL-stimulated RAW 264.7 was used to evaluate inhibitory effect of KSG osteoclast differentiation and gene expression. We counted TRAP (+) multinucleated cells and measured TRAP activity and mRNA expressions of osteoclastogenesis-related genes (NFATc1, MITF, JNK1, cathepsin K, MMP-9) to figure out the effect of KSG on osteoclast. Osteoblastogenesis was also determined in rat calvarial cell. Alkaline phosphatase (ALP) activity, bone matrix protein and collagen synthesis were measured by using murine calvarial cell. Results : KSG inhibited the differentiation of osteoclast precursor cell and expression of genes related osteoclastogenesis like NAFTc1, MITF, c-fos, JNK1, Cathepsin K, MMP-9 and TRAP. KSG increased cell division and function of osteoblast separated from the skull of a rat and ALP synthesis, biosynthesis of bone matrix protein and collagen. Conclusions : Reviewing these results, KSG has efficacy on osteoclast inhibition and osteoblast activation. After further study, KSG will be able to apply for osteoporosis treatment and prevention.