• 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.

• Journal of Life Science
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• v.27 no.3
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• pp.318-324
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• 2017

This study was performed to improve the antioxidant and skin-whitening activities of 70% ethanol extract from Sesamum indicum L. (SIL). The electron-donating ability of the SIL extract was 71.7% at a concentration of $1,000{\mu}g/ml$. The whitening effects that was measured by tyrosinase inhibition assay. As a result, SIL extract was shown 42% at $1,000{\mu}g/ml$ concentration. The cell toxicity on B16F10 melanoma cells of SIL of 70% ethanol extract showed 84.3% at $1,000{\mu}g/ml$ concentration. The microphthalmia-associated transcription factor (MITF), tyrosinase relate protein-1 (TRP-1), tyrosinase relate protein-2 (TRP-2) and Tyrosinase protein and mRNA expression inhibitory effect of SIL extract were measured by western blot and reverse transcription- polymerase chain reaction (PCR) at 50, 250, $500{\mu}g/ml$ concentration. Consequently, the MITF, TRP-1, TRP-2, Tyrosinase protein expression inhibitory effect of SIL extract was decreased by 68.3%, 39.2%, 89.7%, 22.3%, respectively, at $500{\mu}g/ml$ concentration. Moreover, MITF, TRP-1, TRP-2, Tyrosinase mRNA expression inhibitory effect by reverse-transcription-PCR of SIL extract was decreased by 81.8%, 66.5%, 84.2%, 68.1%, respectively, at $500{\mu}g/ml$ concentration. Therefore, we excellently identified the antioxidant activities and whitening effect of SIL extract, and this finding suggested that SIL extract has great potential as a cosmetic ingredients.

• Food Science of Animal Resources
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• v.35 no.5
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• pp.707-713
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• 2015

Royal jelly has been widely used as a health supplement worldwide. However, royal jelly has been implicated in allergic reactions, and we developed a water-soluble royal jelly (WSRJ) without the allergy inducing protein. In this study, we aimed to identify the anti-melanogenic efficacy of WSRJ. B16F1 melanoma cells were first treated with 10 nM α-melanocyte stimulating hormone (α-MSH) and then with various doses of WSRJ. In addition, we investigated the mRNA and protein expression of melanogenesis-related genes such as tyrosinase, tyrosinase related protein-1 (TRP-1) and TRP-2 by reverse transcription-polymerase chain reaction and western blotting. WSRJ directly inhibited tyrosinase and cellular tyrosinase activity, which decreased melanin synthesis in α-MSH stimulated B16F1 melanoma cells a level comparable to that observed with arbutin. WSRJ decreased the mRNA and protein expressions of tyrosinase, TRP-1, and TRP-2, which was comparable to that observed with arbutin. WSRJ has strong anti-melanogenic activity, which invoice direct inhibition of tyrosinase enzyme activity and suppression of expression of melanogenesis related genes. Results from this study suggests that WSRJ is a potential candidate for the treatment of skin pigmentation.

• Korean Journal of Plant Resources
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• v.30 no.1
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• pp.22-28
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• 2017

Sophora japonica has been used for treatment of liver and blood-related diseases in herbology. We evaluated whitening activities of Sophorae Flos and Fructus. Sophorae Flos and Fructus were extracted with methanol (MeOH) and divided into petroleum ether, ethyl acetate(EtOAC) and water fraction. For whitening effect by western blot in B16 F10 cells, we analyzed it by investigating the effect of tyrosinase, TRP-l (tyrosinase-related protein 1), TRP-2 (tyrosinase-related protein 2) and MITF (microphthalmia-associated transcription factor) protein expression. The protein expressions of tyrosinase, TRP-1, TRP-2 and MITF in B16 F10 cells treated with Sophorae Flos and Fructus extract were reduced in a dose-dependant manner. Therefore, these results suggest that Sophorae Flos and Fructus have useful plant resource to be developed as functional cosmetic.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.28 no.1
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• pp.41-52
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• 2015

Objective : This study investigated the inhibitory mechanism of the hypopigmentating effects on ethanol extract of Rosae rugosae Flos (ERR) that has not yet been examined. Methods : We analyzed the anti-melanogenic effects of ethanol extracts from Rosae rugosae Flos by tyrosinase activity, melanin contents. We also examined protein expression levels of tyrosinase, TRP-1, TRP-2, MITF and ERK by western blot analysis in melanoma cells. Results : In this investigation, ERR effectively reduced ${\alpha}$-MSH-stimulated melanin synthesis by suppressing expression of tyrosinase and tyrosinase-related protein-1 (TRP-1). On the other hand, the expression of tyrosinase-related protein-2 (TRP-2) were not affected by treatment with ERR. ERR inhibited the expression of microphthalmia-associated transcription factor (MITF) as a key transcription factor for tyrosinase expression regulating melanogenesis. The upstream signaling pathway including cAMP response element-binding protein (CREB) and MAPKs were also inhibited by ERR. Pretreatment with PD98059, ERK inhibitor, attenuated the inhibitory effect of ERR on ${\alpha}$-MSH-induced tyrosinase activity. Conclusions : Our study suggested that the anti-melanogenic activity of ERR is correlated with the suppression of tyrosinase gene through CREB/MITF/ERK pathway.

• Biomolecules & Therapeutics
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• v.20 no.3
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• pp.340-345
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• 2012

Sarsasapogenin (SAR) is a steroidal sapogenin that is used as starting material for the industrial synthesis of steroids. It has various pharmacological benefits, such as antitumor and antidepressant activities. Since its effect on melanin biosynthesis has not been reported, we used murine melanocyte melan-a cells to investigate whether SAR influences melanogenesis. In this study, SAR significantly increased the melanin content of the melan-a cells from 1 to 10 ${\mu}M$. Based on an enzymatic activity assay using melan-a cell lysate, SAR had no effect on tyrosinase and DOPAchrome tautomerase activities. It also did not affect the protein expression of tyrosinase-related protein 1 and DOPAchrome tautomerase. However, protein levels of tyrosinase and microphthalmia-associated transcription factor were strongly stimulated by treatment with SAR. Therefore, our reports suggest that SAR treatment may induce melanogenesis through the stimulation of tyrosinase and microphthalmia-associated transcription factor expression in melan-a cells.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.37 no.4
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• pp.319-326
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• 2011

In order to investigate the potential of Nigella sativa (N. sativa) oil as an active ingredient for whitening cosmetics, we prepared N. sativa oil. We measured its inhibitory effects on mushroom tyrosinase activity, cellular tyrosinase activity, and melanin synthesis inhibitory activity in B16 melanoma cells. N. sativa oil and its components showed inhibitory activity against mushroom tyrosinase and melanin synthesis. In a melanin synthesis inhibition assay using mouse B16-F10 melanoma cell, it reduced melanin production up to 86 % at a concentration of 10 mg/mL without cytotoxicity. In the study on the melanogenic protein expressions by using RT-PCR and Western blot, N. sativa oil and its components inhibited expression of tyrosinase protein, which is a well-known key protein on melanogenesis, and tyrosinase expression was gradually decreased in a dose-dependent manner. Therefore, this result suggests that N. sativa oil could be used as an active ingredient for whitening cosmetics.

• Journal of Life Science
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• v.23 no.12
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• pp.1445-1453
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• 2013

Human skin is constantly exposed to ultraviolet (UV) radiation, polluted air, and chemical products. UV rays, in particular, will affect the skin in a variety of ways, including causing wrinkles, fine lines, rough skin, and xeroderma, thereby resulting in skin aging. This study aimed to investigate the whitening effects of Juniperus rigida Sieb., which is a cedar tree that is found throughout the world. The whitening efficacy that was measured by tyrosinase inhibition revealed 49.4% efficacy in water extract and 80.0% efficacy in ethanol extract. Among the B16F10 black cells, the effect of the ethanol extract was higher than the effect of the water extract in the restrain creation of melanin pigment, tyrosinase, microphthalmia-associated transcription factor (MITF), tyrosinase related protein-1 (TRP-1), and tyrosinase related protein-2 (TRP-2). Thus, the results of these studies demonstrated that the ethanol extract had greater efficacy than the water extract and Juniperus rigida Sieb. Ethanol extracts could be utilized as materials for functional cosmetics, such as whitening products.

• The Korean Journal of Physiology and Pharmacology
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• v.26 no.2
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• pp.113-123
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• 2022

Diarylpropionitrile (DPN), a selective agonist for estrogen receptor β (ERβ), has been reported to regulate various hormonal responses through activation of ERβ in tissues including the mammary gland and brain. However, the effect of DPN on melanogenesis independent of ERβ has not been studied. The aim of this study is to examine the possibility of anti-melanogenic effect of DPN and its underlying mechanism. Melanin contents and cellular tyrosinase activity assay indicated that DPN inhibited melanin biosynthesis in alpha-melanocyte stimulating hormone-stimulated B16F10 melanoma cell line. However, DPN had no direct influence on in vitro tyrosinase catalytic activity. On the other hand, 17β-estradiol had no effect on inhibition of melanogenesis, suggesting that the DPN-mediated suppression of melanin production was not related with estrogen signaling pathway. Immunoblotting analysis showed that DPN down-regulated the expression of microphthalmia-associated transcription factor (MITF), a central transcription factor of melanogenesis and its down-stream genes including tyrosinase, tyrosinase-related protein (TRP)-1, and TRP-2. Also, DPN attenuated the phosphorylation of protein kinase A (PKA) and cAMP-response element-binding protein (CREB). Additionally, DPN suppressed the melanin synthesis in UVB-irradiated HaCaT conditioned media culture system suggesting that DPN has potential as an anti-melanogenic activity in physiological conditions. Collectively, our data show that DPN inhibits melanogenesis via downregulation of PKA/CREB/MITF signaling pathway.

• YAKHAK HOEJI
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• v.52 no.3
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• pp.165-171
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• 2008

To develop effective skin whitening agents, we tested natural herbal extracts for their melanogenic inhibitory activities. Sedum samentosum was selected for its inhibitory effect on melanogenesis in B16 melanoma cells. Ethanolic extract of S. samentosum (SSE) was evaluated for antioxidative effect and tyrosinase inhibitory activity of melanogenesis. We investigated the changes in protein level and mRNA level of tyrosinase, tyrosinase related protein (TRP)-1 and TRP-2 by using western blotting and RT-PCR, respectively. SSE showed scavenging activities of free radicals and reactive oxygen species (ROS) with the $IC_{50}$ of 342.7 $\mug/ml$ against 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and 64.69 $\mug/ml$ against superoxide radicals in the xanthine/xanthine oxidase system, respectively. SSE treatment suppressed the biosynthesis of melanin up to 46% and reduced tyrosinase activity up to 51% at 100 $\mug/ml$ in B16 melanoma cells. The tyrosinase activity and tyrosinase expression in B16 melanoma cells were reduced in a dose-dependent manner by SSE. Also, SSE was able to significantly inhibit tyrosinase and TRP-1 expression in mRNA level. These results suggest that SSE inhibited melanin production which may be dependent on tyrosinase activity and expression in B16 melanoma cells, and an effective whitening agent for the skin.