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

• Proceedings of the Plant Resources Society of Korea Conference
• /
• 2018.04a
• /
• pp.63-63
• /
• 2018

본 연구에서는 80% 식물성 알코올을 추출 용매로 사용해 오디를 빛을 차단 후 실온에서 3일 간 추출하였다. 3회 여과한 후 최소 온도($40{\sim}60^{\circ}C$)에서 농축한 뒤 동결 건조하여 파우더 형태로 사용하였다. 오디(Morus alba)의 에탄올 추출물은 B16/F10 세포의 항산화 및 멜라닌 합성 억제 효과를 나타내었다. 멜라닌 함량과 세포 내 tyrosinase 활성을 Western blotting으로 측정 하였다. Tyrosinase와 tyrosinase-related protein (TRP) -1은 tyrosinase-related protein (TRP) -2보다 강력하게 억제되었으며, 이들 결과는 tyrosinase와 TRP-1은 흑갈색을 띠는 eumelanin의 생합성의 억제와 강한 상관관계가 있음을 보여 주었다. ${\alpha}$-melanocyte-stimulating hormone (${\alpha}$-MSH) 처리 한 B16/F10 흑색 종 세포에서 M. alba 에탄올 추출물은 멜라닌 생성 연관 단백질의 발현 및 멜라닌 생성이 용량 의존적으로 억제 하였다. 멜라닌 함량과 세포 내 tyrosinase 활성을 Western blotting으로 측정 하였다. 또한 DPPH와 SOD를 사용하여 항산화 활성을 분석하였고 총 폴리 페놀과 총 플라보노이드 함량을 측정 하였다. MTT assay 분석을 사용하여 M. alba 에탄올 추출물의 세포 독성을 측정 하였다. B16/F10 멜라닌 생성 세포의 tyrosinase 저해 활성 및 사멸 효과가 일반적으로 효과적이었다. 따라서 M. alba 에탄올 추출물은 항산화 및 미백 효과를 나타내며, 기능성 화장품의 천연 성분으로서 우수한 것으로 여겨진다.

• Journal of Photoscience
• /
• v.9 no.2
• /
• pp.209-212
• /
• 2002

Cutaneous hyperpigmentation is a well-known consequence of both acute and chronic X-irradiation, although the molecular mechanisms involved are not well understood. Recently, protein kinase C-$\beta$ (PKC-$\beta$) was shown to activate tyrosinase, a key and the rate-limiting enzyme in melanogenesis [1]. In this study, we have investigated its role in mediating ionizing radiation-induced pigmentation by exposing cultured human melanocytes to X-irradiation. Increased tyrosinase activity after the 4 Gys exposure was observed within 48 hrs and total melanin content doubled after 7 days. Interestingly, tyrosinase mRNA level was not affected by X-irradiation. However, there was a 2-3 fold increase in PKC-$\beta$ mRNA after 48 hours of irradiation, coinciding with the increase in tyrosinase activity. This induction was not due to non-specific heat generated during the irradiation because when melanocytes were incubated at 4$0^{\circ}C$, there was no induction of PKC-$\beta$ mRNA. Taken together, these data suggest that X-irradiation induces cutaneous hyperpigmentation, at least in part, by up-regulating the level of PKC-$\beta$.

• Natural Product Sciences
• /
• v.20 no.1
• /
• pp.33-38
• /
• 2014

In recent times, the demand for edible medication for the treatment of hyperpigmentation has increased significantly. Therefore, the discovery of a stable, safe and inexpansive antimelanogenic component from natural substances, such as grains, is of particular interest. The levels and activities of some metabolites and/or enzymes can be increased. In the present study, we investigated the antimelanogenic effects of water-soluble extracts from barley (BE), malt (ME) and germinated barley (GBE) in melan-a cells. The inhibitory effects of ME and GBE on melanin production were significantly greater than that of BE. Interestingly, the content of ferulic acid, the proposed active component of barley, was also higher in ME and GBE than in BE by HPLC analysis. Western blot analysis of the expression of melanogenic enzymes in melan-a cells treated with BE, ME or GBE indicated the expression of both tyrosinase and tyrosinase-related protein 2 (TRP-2) significantly decreased after treatment with BE, ME or GBE. These results suggest that besides BE, ME and GBE also inhibit melanin production most likely through suppression of tyrosinase and TRP-2 expression. ME and GBE were more efficacious at inhibiting melanin production than BE was and may also represent potential skin-whitening agents.

• YAKHAK HOEJI
• /
• v.49 no.5
• /
• pp.375-379
• /
• 2005

The green marine algae, Capsosiphon fulvescens is one of the important economic seaweeds cultured in Korea. In this study, we investigated the effects of Capsosiphon fulvescens on melanogenesis using B16 cells. Our results showed that Capsosiphon fulvescens significantly inhibits melanin synthesis and it reduces the activity of tyrosinase, the rate-limiting melanogenic enzyme. Western Blot analysis using anti-tyrosinase antibody revealed that Capsosiphon fulvescens ($10\~40\;{\mu}g/ml$ decreased tyrosinase protein levels. Cell proliferation was dose-dependently inhibited by 10, 20 and 40 ${\mu}g/ml$ Cap­sosiphon fulvescens, without cytotoxicity and morphological change. These results suggest that the depigmenting effect of Capsosiphon fulvescens is correlated with the suppression of tyrosinase activity and protein level, which are key enzymes for melanogenesis.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.22 no.2
• /
• pp.371-376
• /
• 2008

In this study, we have investigated the hypo-pigmentary mechanism of methanol extract of Forsythiae Fructus in human melanocyte cell line, HM3KO. Treatment of HM3KO cells with Forsythiae Fructus extract markedly inhibited melanin biosynthesis in a dose-dependent manner. Decreased melanin contents occurred through the decrease of tyrosinase protein and activity. The mRNA levels of tyrosinase and tyrosinase-related protein 1 (TRP-1) were also reduced by Forsythiae Fructus extract. Moreover, the level of intracellular cyclic AMP (cAMP) was significantly decreased by treatment of Forsythiae Fructus extract. These results suggest that Forsythiae Fructus reduces melanin synthesis by down regulation of tyrosinase mRNA transcription, and this is closely related to the cAMP-dependent pathway.

• Journal of the Society of Cosmetic Scientists of Korea
• /
• v.48 no.1
• /
• pp.11-24
• /
• 2022

It has been reported that the ethanolic extract of the root of Piper methysticum (P. methysticum) inhibits melanogenesis in melanocyte stimulating hormone (MSH)-activated B16 melanoma cells. Flavokawain B (FKB) and Flavokawain C (FKC) isolated from this extract have been found to inhibit melanin production based on anti-melanogenesis activity. This study was designed to find out the inhibition and its process of FKB and FKC on melanin synthesis in melan-a melanocytes. FKB and FKC inhibited melanogenesis at 10 μM, 5 μM respectively in melan-a melanocytes. However, they did not inhibit extracellular tyrosinase activity from melan-a melanocytes. FKB reduced the protein level of tyrosinase (Tyr), tyrosinase-related protein 1 (TRP-1), tyrosinase-related protein 2 (TRP-2), microphthalmia-associated transcription factor (MITF) and the mRNA level of Tyr and TRP-1. FKC reduced the protein level of TRP-2 and MITF and the mRNA level of TRP-1 and Tyr. The reduced expression of Tyr and TRP-1 might be resulted from the decreased MITF which regulates major melanogenic proteins. However, since the mRNA expression of MITF did not change by FKB and FKC treatment, the effects of FKB and FKC on extracellular signal regulating kinase (ERK)/AKT phosphorylation, known to regulate the degradation of MITF, were confirmed. FKB and FKC significantly increased the phosphorylation of ERK1/2, not in AKT. These results suggest that FKB and FKC may be helpful as a potential depigmenting agent for various hyper-pigmentary disorders.

• Korean Journal of Pharmacognosy
• /
• v.45 no.3
• /
• pp.262-267
• /
• 2014

Royal jelly composes of many components, especially protein. Protein is a major factor which cause allergy. We focused on water soluble royal jelly (WSRJ) that was removed allergy - inducing protein. 10-hyroxy-2-decenoic acid content of WSRJ is 2.42 g/100 g, which is double compared to that of lypophilized RJ. To further access WSRJ as a cosmetic ingredient and potential external treatment for topical use, we investigated its ability to inhibit tyrosinase activity and melanin biosynthesis on melanogenesis in B16F1 melanoma cells. We found that WSRJ increased the cell viability in B16F1 melanoma cell and WSRJ (1~10 mg/ml) inhibited melanin synthesis in with 10 nM ${\alpha}$-melanocyte-stimulating hormone (${\alpha}$-MSH) for 48 h. WSRJ inhibited direct tyrosinase activity, which decreased melanin synthesis in ${\alpha}$-MSH stimulated B16F1 melanoma cells. Thease findings suggest that WSRJ induces the down regulation of melanogenesis by inhibiting tyrosinase activation.

• Microbiology and Biotechnology Letters
• /
• v.19 no.3
• /
• pp.281-289
• /
• 1991

Neurospora crassa (KCTC 6079) produces tyrosinase (EC 1.14.18.1) during sexual differentiation under derepressed conditions in the presence of inducers such as amino acid analogues, antimetabolites or protein synthesis inhibitors. The selection of inducer concentration and induction time as well as inducer type are critical for the optimization of the enzyme production. The best inducer was found to be cycloheximide. Since cycloheximide was toxic to the cells, an optimal inducer concentration and an optimal induction time were determined to maximize the enzyme production from batch cultures. Mathematical models for the cell growth and the enzyme production were proposed and used for process optimization. By optimizing the induction conditions, maximum tyrosinase productivity was increased significantly.

• Journal of Microbiology and Biotechnology
• /
• v.28 no.12
• /
• pp.2121-2132
• /
• 2018

Abnormal melanin synthesis results in several hyperpigmentary disorders such as freckles, melanoderma, age spots, and other related conditions. In this study, we investigated the anti-melanogenic effects of an extract from the microalgae Chlamydomonas reinhardtii (CE) and potential mechanisms responsible for its inhibitory effect in B16F10, normal human epidermal melanocyte cells, and human skin-equivalent models. The CE extract showed significant dose-dependent inhibitory effects on ${\alpha}$-melanocyte-stimulating, hormone-induced melanin synthesis in cells. Additionally, the CE extract exhibited suppressive effects on the mRNA and protein expression of microphthalmia-associated transcription factor, tyrosinase, tyrosinase-related protein-1, and tyrosinase-related protein-2. The CE extract also inhibited the phosphorylation of protein kinase A and extracellular signal-related kinase, which function as upstream regulators of melanogenesis. Using a three-dimensional, reconstructed pigmented epidermis model, the CE-mediated, anti-pigmentation effects were confirmed by Fontana-Masson staining and melanin content assays. Taken together, CE extract can be used as an anti-pigmentation agent.