• Journal of Physiology & Pathology in Korean Medicine
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• v.22 no.2
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• pp.346-349
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• 2008

To investigate whether aqueous extract from Aconitum koreanum (AEAK) effects in the process of melanin synthesis, the expression of tyrosinase-related proteins (TRPs) by immunohistochemical methods were performed in ultraviolet B (UVB) irradiated skin of guinea pig. The irradiation of UVB (60 mJ/day) was performed for 3 days and treated with AEAK for 15 days. About the color evaluation, the visual scores of UV B irradiated guinea pig with AEAK treatment were slightly lower than those in the UV B alone irradiated ones. At day 15 after UVB exposure, immunohistochemical analysis for TRPs expression were performed. The intensive expression of tyrosinase was mainly observed over epidermis with skin appendage and in the cells of dermis. Slight increase of these reaction was induced in response to UVB in the spinous and granular layer of epidermis, but similar expression in the AEAK treated guinea pig as normal one. The TRP-1 and TRP-2 expression were not detected in the skin of normal guinea pig. But intensive expression for TRP-1 and TRP-2, especially TRP-2, induced by UV B irradiation in the cells of dermis. These expressions were decreased in the AEAK treated guniea pig. Collectively, these results suggest that AEAK has a potential to inhibit synthesis through regulation of TRPs expression in the skin of guinea pig, but better understanding the function of AEAK, more research should be done in the effects of AEAK on the function of TRPs in melanogesis.

• Korean Journal of Pharmacognosy
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• v.51 no.3
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• pp.171-177
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• 2020

The purpose of this study was to investigate the anti-melanogenic effects of the extracts from Decaschistia intermedia craib (EDI). In this study, we examined the effects of EDI on mushroom tyrosinase activity in in vitro, melanin contents, and expression levels of mRNA and proteins of melanogenesis-related genes in B16F10 melanoma cells. The treatment of EDI significantly decreased both tyrosinase activity and melanin contents in B16F10 cells with dose-dependent manner. In addition, we found that the expression of mRNA or proteins of melanogenic proteins, such as, a-melanocyte-stimulating hormone (a-MSH)-induced microphthalmia associated transcription factor (MITF), tyrosinase, tyrosinase related protein-1 (TRP-1), and TRP-2 was significantly downregulated with dose-dependent manner in the EDI-treated B16F10 cells compared to controls. Our results suggest that the EDI inhibits cellular melanogenesis through downregulation of a-MSH-stimulated melanin synthesis. Thus EDI may potentially be an effective whitening agent.

• Journal of Life Science
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• v.24 no.5
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• pp.485-490
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• 2014

Poria cocas has been reported to be effective in skin whitening. However, the direct effect of P. cocas ethanol extracts (PCEE) on melanin synthesis has not been scientifically studied. To elucidate the direct effect of PCEE on melanogenesis, a 3,4-dihydroxyindole-2-carboxylic acid (DOPA) synthesis assay, tyrosinase activity assay, and Western blotting for melanogenic proteins, including tyrosinase, tyrosinase-related protein (TRP)-1, and TRP-2 were performed in mouse B16F1 cells. The results revealed that PGEE inhibited melanin production in a dose-dependent manner by blocking the synthesis of DOPA. Although the activation of tyrosinase was not affected, the expression levels of TRP-1 and TRP-2 were controlled. These results suggest that PCEE has a whitening effect, indicating that it may be a useful agent in the development of whitening cosmetics.

• Journal of Applied Biological Chemistry
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• v.58 no.4
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• pp.333-338
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• 2015

The objective of this study was to investigate the whitening effect of Salvia miltorrhiza Bunge water extract (SM-W) in human epidermal melanocyte (HEM). Mushroom tyrosinase inhibitory effect of SM-W was approximately 42% at $1,000{\mu}g/mL$. The HEM cellular tyrosinase and melanin synthesis inhibition activity were 26 and 25% at $5{\mu}g/mL$, respectively. Whitening related proteins and mRNAs including tyrosinase, tyrosinase related protein 1 (TRP-1) and TRP-2, and microphthalmia associated transcription factor were reduced by SM-W treatment. In addition, the cAMP expression inhibitory effect of SM-W was decreased by 41% at $5{\mu}g/mL$ concentration. These results indicated that Salvia miltorrhiza Bunge could be used to the possible utilization of functional cosmetic ingredients by confirming whitening activity related with melanin content.

• Biomolecules & Therapeutics
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• v.22 no.1
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• pp.35-40
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• 2014

Resveratrol is a polyphenolic compound found in various natural products such as grapes and berries and possesses anti-cancer, anti-hyperlipidemia, and anti-aging properties. Recently, it has been reported that resveratrol inhibits ${\alpha}$-melanocyte-stimulating hormone signaling, viability, and migration in melanoma cells. However, these effects have not been confirmed in vivo, specifically brownish guinea pigs. To evaluate the potential of resveratrol as a regulator of melanin for hyperpigmentation therapy, the influence of resveratrol on pigmentation was investigated by ultraviolet B-induced hyperpigmentation in brownish guinea pig skin. We found that resveratrol reduced the expression of melanogenesis-related proteins tyrosinase, tyrosinase-related proteins 1 and 2, and microphthalmia-associated transcription factor in melanoma cells. Furthermore, topical application of resveratrol was demonstrated to significantly decrease hyperpigmentation on ultraviolet B-stimulated guinea pig skin in vivo. Based on our histological data, resveratrol inhibits melanin synthesis via a reduction in tyrosinase-related protein 2 among the melanogenic enzymes. This study is the first to provide evidence supporting resveratrol as a depigmentation agent, along with further clinical investigation of resveratrol in ultraviolet B-induced skin disorders such as hyperpigmentation and skin photoaging.

• Korean Journal of Medicinal Crop Science
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• v.19 no.3
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• pp.177-184
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• 2011

This study was performed to assess the antioxidant activities and whitening effects of Agrimonia pilosa Ledeb on melanin synthesis. The whitening effects of Agrimonia pilosa Ledeb water extracts were examined by in vitro mushroom tyrosinase assay and B16BL6 melanoma cells. We assessed inhibitory effect of Agrimonia pilosa Ledeb water extract on expression of melanogenic enzyme proteins including tyrosinase, tyrosinase-related protein 1 (TRP-1) and tyrosinase-related protein 2 (TRP-2) in B16BL6 cells. Inhibitory effect of Agrimonia pilosa Ledeb onto free radical generation was determined by measuring DPPH and hydroxyl radical scavenging activitie. Our results indicated that Agrimonia pilosa Ledeb water extract effectively inhibited free radical generation. In DPPH and hydroxy radical scavenging activity, Agrimonia pilosa Ledeb water extract had a potent anti-oxidant activity in a dose-dependent manner. They significantly inhibited tyrosinase activity in vitro and in B16BL6 melanoma cells. Also, Agrimonia pilosa Ledeb suppressed the expression of tyrosinase in B16BL6 melanoma cells. These results show that Agrimonia pilosa Ledeb inhibited melanin production on the melanogenesis. The underlying mechanism of Agrimonia pilosa Ledeb on whitening activity may be due to the inhibition of tyrosinase activity. We suggest that Agrimonia pilosa Ledeb may be useful as new natural active ingredients for antioxidant and whitening cosmetics.

• KSBB Journal
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• v.28 no.2
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• pp.137-145
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• 2013

In order to develop new skin whitening agents, we prepared the $CH_2Cl_2$ layer (NGC) and BuOH layer (NGB) of 75% EtOH extract of the Nelumbinis nucifera Gaertner. We measured their tyrosinase inhibitory activity in vitro and melanin synthesis inhibitory activity in B16-F1 melanoma cells. They did not show inhibitory activity against mushroom tyrosinase but showed melanin synthesis inhibitory activity in a dose-dependent manner. In a melanin synthesis inhibition assay, NGC and NGB suppressed melanin production up to 52% and 46% at a concentration of $100{\mu}g/mL$, respectively. To elucidate the mechanism of the inhibitory effects of NGC and NGB on melanogenesis, we measured the expression of melanogenesis-related proteins by western blot assay. As a result, NGC suppressed the expression of tyrosinase, tyrosinase related protein 1 (TRP-1), tyrosinase related protein 2 (TRP-2), phosphorylated cAMP responsive element binding (p-CREB) protein, and microphthalmia associated transcription factor (MITF). And NGB inhibited the protein expression of tyrosinase and MITF, but had no significant effect on TRP-1, TRP-2, and p-CREB expression. Moreover, NGB increased the expression of phosphorylated extracellular signal-regulated kinase (p-ERK). In addition, we examined the inhibitory effect on the glycosylation of tyrosinase. As a result, NGC and NGB inhibited the activity of ${\alpha}$-glucosidase in vitro and the glycosylation of tyrosinase in B16-F1 melanoma cells. From these results, we concluded that NGC and NGB could be used as active ingredients for skin whitening.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2023.04a
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• pp.56-56
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• 2023

The cone of Pinus rigida × Pinus taeda (PRT), a plant in the Pinaceae family, has long been used in traditional medicine to treat hemostasis, bruises, and burns. Previous research has shown that regulating oxidation-reduction reactions in reactive oxygen species can help inhibit melanogenesis, the process of melanin synthesis, which is a common target for addressing hyperpigmentation. Inhibiting tyrosinase is also known to be effective in this regard. Based on these findings, we conducted an investigation into the inhibitory effect of the ethyl acetate fraction of PRT (ERT) on melanogenesis in B16 F10 cells. We know that the expression levels of melanin biosynthesis-related proteins, including tyrosinase, TRP-1, and TRP-2, are regulated by MITF (microphthalmia-associated transcription factor) and cAMP, with cAMP affecting the activity of protein kinase A (PKA). PKA can reduce melanogenesis, and CREB reduces the phosphorylation of melanin-producing enzymes. In addition, the MAPK signaling pathway, composed of ERK, JNK, p38, and other factors, is also known to play a role in the inhibition of melanogenesis in melanocytes. Our immunoblotting results showed that ERT inhibited the expression of melanin production-related proteins (tyrosinase, TRP-1, TRP-2, and MITF) that were significantly increased by a-MSH treatment to promote melanin production. Furthermore, the phosphorylation levels of factors related to cAMP/PKA/CREB and MAPK signaling pathways were significantly reduced without affecting the total form. In conclusion, we believe that treatment with ERT can inhibit melanin synthesis by modulating the phosphorylation of cAMP/PKA/CREB and MAPK signaling pathways at the cellular level. These findings suggest the potential of ERT as a raw material for functional cosmetics and pharmaceuticals, thanks to its antioxidant activity and ability to inhibit melanogenesis. We thought that these findings of ERT as a natural plant resource will inspire further research and development in this area.

• Proceedings of the Korean Society of Applied Pharmacology
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• 2007.11a
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• pp.129-139
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• 2007

Many modalities of treatment for acquired skin hyperpigmentation are available including chemical agents or physical therapies, but none are completely satisfactory. The ideal depigmenting compound should have a potent. rapid and selective bleaching effect on hyperactivated melanocytes, carry no short- or long-term side-effects and lead to a permanent removal of undesired pigment. acting at one or more steps of the pigmentation process. Depigmentation can be achieved by regulating (i) the transcription and activity of tyrosinase, tyrosinase related protein-1 (TRP-1), tyrosinase related protein-2 (TRP-2), and/or peroxidase; (ii) the uptake and distribution of melanosomes in recipient keratinocytes and (iii) melanin and melanosome degradation and turnover of pigmented keratinocytes. One of the interesting point for development of skin whitening agent is Mitf(Microphthalmia-associated transcription factor). Mitf belongs to the basic helix-loop-helix-zip family of trabscription factors and it is crucial as it regulates both melanocyte proliferation as well as melanogenesis and is the major regulator of tyrosinase and the related enzymes (TRPs), as well as many melanosome structural proteins such as pMel17. Recently, we developed MITF-down-regulating agents from natural and synthetic sources, which have anti-melanogenic effect on in vitro and in vivo. We suggested that potent MITF-down regulating agents might be used for skin whitening cosmeceuticals.

• Journal of Life Science
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• v.29 no.11
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• pp.1192-1199
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• 2019

The intention of this study was to confirm the possible use of an ethanol extracts of Nelumbinis Rhizomatis Nodus (NRN) as a cosmetic material. To this end, we extracted NRN with 70% ethanol and performed biological activity evaluation of whitening efficacy and wrinkle reduction. We performed cellular tyrosinase inhibition and melanin contents assay to check the whitening activity of NRN and carried out a toxicity evaluation of NRN via an MTT assay and the amounts of associated proteins that affect melanin production in a melanoma cell line (B16F10). And collagenase inhibitory assay was performed for the evaluation of anti-wrinkle of samples. In addition, a toxicity evaluation using an MTT assay and matrix metalloprotease (MMP-1) and procollagen synthesis inhibition by NRN were evaluated in a fibroblast cell line (CCD-986sk). Western blot results for the whitening activity evaluation revealed that the levels of two proteins related to melanin production, tyrosinase-related protein-1 (TRP-1) and tyrosinase-related protein-2 (TRP-2), were decreased in a dose-dependent manner. Moreover, collagenase inhibition activity at a concentration of $500{\mu}g/ml$ NRN by measuring epigallocatechin-3-gallate (EGCG) was increased by more than 80% compared to the control group. Meanwhile, procollagen synthesis was reduced by 68.8% in the UVB-induced CCD- 986sk cells group whereas collagen synthesis recovered by 80.2% with $25{\mu}g/ml$ NRN. The MMP-1 expression rate showed 20.2% reduction at $25{\mu}g/ml$. The results of the experiments verified the whitening and wrinkle suppression effects of NRN and confirmed that it could be used as a safe natural cosmetic material in the future.