• Journal of the Korean Society of Food Science and Nutrition
• /
• v.43 no.5
• /
• pp.705-711
• /
• 2014

In order to develop a skin-whitening agent, melanin contents and intracellular tyrosinase activity were determined by western blotting. Ethyl acetate fractions of 80% ethanol extracts from lily (Lilium Oriental Hybrid 'Siberia') bulbs (R-EA) inhibited melanin synthesis in a dose-dependent manner in ${\alpha}$-melanocyte stimulating hormone (${\alpha}$-MSH)-treated B16/F10 murine melanoma cells. Intracellular tyrosinase activity and melanin contents were suppressed by 45% and 74%, respectively, in response to treatment with $100{\mu}g/mL$ of R-EA. Examination of protein expression associated with ${\alpha}$-MSH-induced melanogenesis showed that tyrosinase related protein (TRP)-1 was inhibited more strongly than tyrosinase, and these results were correlated with stronger inhibition of melanin synthesis than intracellular tyrosinase activity. Taken together, R-EA containing p-coumaric acid and resveratrol could be used as a hypopigmentation agent through suppression of sustained extracellular signal-regulated kinase (ERK) activation via melanogenic induction.

• Journal of the Society of Cosmetic Scientists of Korea
• /
• v.35 no.3
• /
• pp.243-250
• /
• 2009

To develop a new natural whitening agent for cosmetics, we investigated the inhibitory effects of Poria cocos Bark extracts (PCBE) and its active compound on melanogenesis. PCBE showed ROS scavenging activities in 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and xanthine/xanthine oxidase system with the $IC_{50}$ values of $19.4{\pm}2.21{\mu}g$/mL and $IC_{50}=103{\pm}3.33{\mu}g$/mL, respectively. PCBE reduced intracellular tyrosinase activity about 34 % at concentration of $50{\mu}g$/mL. And PCBE reduced melanin contents of B16 melanoma cells about 51 % at concentration of $50{\mu}g$/mL without cell cytotoxicity (below $100{\mu}g$/mL). We purified one active compound from PCBE and identified its structure. It was identified as 3-$\beta$-hydroxylanosta-7,9(11),24-trien-4-oic acid, triterpene family, by $^1H$-NMR, $^{13}C$-NMR and Mass analysis. 3-$\beta$-hydroxylanosta-7,9(11),24-trien-4-oic acid showed ROS scavenging activities in DPPH radical and xanthine/xanthine oxidase system with the $IC_{50}$ values of $4.3{\pm}0.15{\mu}g$/mL and $54{\pm}1.67{\mu}g$/mL, respectively. Also, it was shown that 3-$\beta$-hydroxylanosta-7,9(11),24-trien-4-oic acid reduced intracellular tyrosinase activity about 43 % at concentration of $10{\mu}g$/mL. And it inhibited melanin synthesis in a dose dependent manner ($IC_{50}=3.6{\mu}g$/mL) without cell cytotoxicity (below $100{\mu}g$/mL). 3-$\beta$-hydroxylanosta-7,9(11),24-trien-4-oic acid inhibited tyrosinase, TRP-1 and TRP-2 expression at protein level. These results suggest that PCBE and 3-$\beta$-hydroxylanosta-7,9(11),24-trien-4-oic acid reduced melanin formation by the inhibition of tyrosinase activity and expression in B16 melanoma cells. Therefore, we suggest that PCBE could be used as a useful whitening agent.

• Journal of Life Science
• /
• v.32 no.3
• /
• pp.222-231
• /
• 2022

This study aimed to confirm the possibility of being used as a cosmetic material material through the verification of the whitening and anti-inflammatory activities of an ethyl acetate fraction from Glechoma hederacea var. longituba (GHEA). The observed electron donating and ABTS⁺ radical scavenging abilities of GHEA were 89.6% and 88.7%, respectively, at 1,000 ㎍/ml concentration, with a tyrosinase inhibitory effect of 22.3% at the same concentration. For cell viability, a rate of 80% or more was observed in all concentrations that treated GHEA on melanoma and macrophage cells. Protein and mRNA expression inhibition was measured by Western blot and RT-PCR for 25, 50, and 100 ㎍/ ml concentrations, and it was confirmed that expression decreases in a concentration-dependent manner as GHEA concentration increases. The inhibition of the whitening-related factors MITF and TRP-2 were superior following GHEA treatment than those of the control group treated with kojic acid of 100 ㎍/ml concentration. For tyrosinase, the lowest mRNA expression rate was 29.1% at 100 ㎍/ml which confirmed excellent inhibition. In analyzing the effects of pro-inflammatory cytokines IL-1β, IL-6, and TNF-α on protein and mRNA expression, IL-6 and TNF-α showed high protein and mRNA inhibition compared to a vitamin C control group. Based on these experimental results, GHEA could be applied as a natural cosmetic material.

• The Journal of Korean Medicine
• /
• v.38 no.4
• /
• pp.62-81
• /
• 2017

Objectives: As interests in the beauty of skin is growing continuously, more people are focusing on white and clean skin. Melanin is the major factor that determines skin color. The abnormal concentration of melanin causes various skin diseases such as vitiligo, freckles, and melasma. This study investigated the inhibitory effect of Eriobotryae Folium extracts (EF) with phreatic water (PW) on the melanin synthesis. Methods: The effect of EF on melanin synthesis was evaluated by using mouse melanoma cells (B16F10). To define the mechanisms, real-time PCR and western blot were used. We also evaluated the inhibitory effects of EF and PW on melanin synthesis by using HRM-2 melanin-possessing hairless mice. After UVB irradiation, melanin differences between the skin parts that were treated and untreated with EF and PW. Levels of mRNA were measured by real-time quantitative PCR and histological analysis of the dorsal skin was conducted by hematoxylin and eosin staining. Results: EF inhibited various mechanisms of melanogenesis, and the effect was increased when combined with PW. In vitro experiments have shown that EF inhibited the expressions of tyrosinase related protein-1 (TRP-1) mRNA, tyrosinase mRNA, microphthalmia-associated transcription factor (MITF) mRNA and the tyrosinase inhibitory activation, but it stimulated the extracellular regulated kinase (ERK) mRNA expression. In vivo experiments have shown that EF prevented melanogenesis in the mice dorsal skin and inhibited TRP-1 mRNA expression. Also these effects were increased when combined with PW. Conclusions: EF and PW might be a new and effective treatment for whitening and treating pigmentation of skin.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.42 no.4
• /
• pp.550-555
• /
• 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 Applied Biological Chemistry
• /
• v.65 no.2
• /
• pp.93-99
• /
• 2022

Lysosome organelle extract (LOE) was derived from egg whites. The lysosome is an intracellular organelle that contains several hydrolysis enzymes. Previous studies have reported that LOE performs important functions, such as melanin de-colorization and anti-melanin production in B16F10 melanoma cells. However, its principal molecular and cellular mechanisms have not been elucidated till date. In non-cytotoxic conditions, LOE significantly inhibited α-MSH induced melanin synthesis of murine B16F10 cells. The anti-melanogenic activity of LOE was mediated by suppressing the mRNA expression of tyrosinase enzyme, tyrosinase related protein-1/2 (TRP-1/2), and microphthalmia-associated transcription factor (MITF) genes. By performing western blot analysis, we found that LOE significantly attenuated melanogenesis. In this case, LOE helped in increasing extracellular receptor kinase (ERK) phosphorylation in α-MSH induced B16F10 cells. Furthermore, MITF is found to be a key regulatory transcription factor in melanin synthesis; it was down-regulated by LOE through ERK phosphorylation. In this experiment, PD98059 (MEK inhibitor) was used to check whether LOE directly regulated the activity of ERK. Although LOE exerted inhibitory effect on melanin synthesis, we could not observe this effect in PD98059-treated α-MSH induced B16F10. These results strongly indicate that LOE is related to ERK activation and MITF degradation in anti-skin pigmentation. Hence, LOE should be utilized as a whitening agent of skin in the near future.

• Journal of Applied Biological Chemistry
• /
• v.65 no.3
• /
• pp.231-237
• /
• 2022

Biorenovation is a method that converts existing compounds into new compounds through the enzymatic action of microorganisms. Biorenovation has expected effects such as reducing toxicity of compounds and increasing their activity. In this study, we successfully synthesized Luteolin-7-O-glucoside (L7G) through biorenovation and investigated its inhibitory effect on melanin production in α-Melanocyte stimulating hormone induced B16F10 mouse melanoma cells. We confirmed that Luteolin was toxic at 50, 100 and 200 µM, but our L7G in same concentration was not toxic for B16F10 mouse melanoma cells and also showed significant reduction in melanin production and tyrosinase activity. In addition, while investigating the effect of L7G on factors involved in melanin synthesis through western blotting, we were able to confirm that the MITF and tyrosinase protein synthesis was inhibited in treatment with L7G, however, tyrosinase related protein-1 (TRP-1) and dopachrome tautomerase (TRP-2) expression was not affected. So we derived a conclusion that through biorenovation we could produce compounds like L7G with improved activity and reduced toxicity for possible use as an active ingredient with whitening functionality in cosmetics.It also suggests that the application of biorenovation has potential usefulness in developing anti-inflammatory materials. It also suggests that the application of bio-renovation has potential usefulness in the development of inflammatory material. We applied Biorenovation technology to Distylium racemosum extract (DR) to generate Distylium racemosum biorenovation product (DRB), and investigated the anti-inflammatory properties of DRB in lipopolysaccharide (LPS)-treated RAW264.7 macrophages. We are applying technology to Biorenovation Distylium racemosum extract (DR) Distylium racemosum was to create a biorenovation product (DRB), lipopolysaccharide (LPS) investigated the anti-inflammatory properties of DRB in RAW264.7 macrophages treated for.

• Journal of Life Science
• /
• v.28 no.7
• /
• pp.802-810
• /
• 2018

Aging is accompanied by changes in the body, such as graying hair, wrinkles, and black spots composed of lipid peroxides and proteins. Melanin is a polymer substance produced by an oxidation polymerization reaction from tyrosine, and it determines the color of hair and skin. It has been reported that melanin is synthesized by melanocyte, and its excessive production by reactive oxygen species is associated with aging. The purpose of this study was to determine the direct effects of Musa paradisiaca peel ethanolic extract (MPEE) on antioxidative activity and melanin synthesis. It was observed that the antioxidant activity of MPEE was similar to that of vitamin C, a positive control, in both DPPH radical scavenging assay and reducing power assay. In order to examine cytotoxicity prior to cell experimentation, 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was performed for B16F1 cells. MPEE was not cytotoxic at $32{\mu}g/ml$ or less. In addition, MPEE increased melanin synthesis in live cells in addition to tyrosinase activity and melanin synthesis in dihydroxyphenylalanine (DOPA)-oxidation assay in vitro. Moreover, MPEE increased melanin synthesis in cells aged by pretreatment with $H_2O_2$. The expression levels of tyrosinase-related protein (TRP)-1, TRP-2, and superoxide dismutase (SOD)-2 by western blot analysis were increased in the presence of MPEE. These results suggest that MPEE could promote the melanin synthesis as an antioxidative substance.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.46 no.1
• /
• pp.34-38
• /
• 2017

Myelophycus simplex Papenfuss, a type of brown algae, is known to be majorly distributed in along the southern coast of Korea and Japan. The purpose of this study was to investigate the effects of M. simplex Papenfuss methanol extract (MSPME) on melanogenesis in ${\alpha}$-melanocyte-stimulating hormone-stimulated B16F10 melanoma cells. Melanin contents of B16F10 melanoma cells were decreased by 27, 41, and 59% in a dose-dependent manner, upon MSPME treatment at 100, 300, and $500{\mu}g/mL$, respectively. Tyrosinase activities in B16F10 melanoma cells were decreased by 18, 49, and 61% in a dose-dependent manner, upon MSPME treatment at 100, 300, and $500{\mu}g/mL$, respectively. MSPME suppressed expression of tyrosinase, tyrosinase-related protein-1, tyrosinase-related protein-2, and melanocyte-inducing transcription factor in B16F10 melanoma cells. Concentration of $50{\mu}g/mL$ of MSPME especially induced greater decreases in tyrosinase activity, melanin contents, and melanogenic enzyme protein expressions. This results indicate that MSPME inhibits melanin synthesis and tyrosinase activity, and M. simplex Papenfuss extract may be an ideal candidate as a skin whitening agent.

• The Korea Journal of Herbology
• /
• v.27 no.4
• /
• pp.73-80
• /
• 2012

Objectives : Under normal condition melanin protects the skin from extracellular stimuli including ultraviolet (UV)-induced oxidative skin damages, but excess production and accumulation of melanin can induce hyperpigmentation causing esthetic problems. Therefore, in this study we tried to search for natural skin whitening materials from marine natural resources. Methods : Water and ethanol extracts of marine natural resources were prepared from Porphyra thalli (PT), Laminariae thallus (LT), Ostreae concha (OC), Sargassum thallus (ST), Undaria thallus (UT), Codium thalli (CT), Enteromorpha thalli (ET), Syngnathoides biaculeatus (SB), and Hippocampus coronatus (Hc). Their effects against UVB and ${\alpha}$-melanocyte stimulating hormone (${\alpha}$-MSH)-induced melanogenesis were investigated based on melanin formation in B16 mouse melanoma cells. The mRNA and protein expression of enzymes involved in the melanogenic process were further examined by reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blot analysis, respectively. Results : Water extract of Ostreae concha (OCW/E) effectively inhibited UVB and ${\alpha}$-MSH-induced melanin production in B16 melanocytes, which seemed to be mediated by inhibition of mRNA expression of tyrosinase and tyrosinase-related protein 1 (TRP-1). In another experiment, ethanol extracts from Porphyra thalli (PTE/E), Laminariae thallus (LTE/E), Sargassum thallus (STE/E), Undaria thallus (UTE/E), Codium thalli (CTE/E), Syngnathoides biaculeatus (SBE/E), and Hippocampus coronatus (HcE/E) significantly suppressed UVB and ${\alpha}$-MSH-induced melanin formation. Furthermore, ethylacetate fraction isolated form LTE/E (LTE/EEt) decreased UVB and ${\alpha}$-MSH-elevated extracellular melanin levels via inhibition of tyrosinase protein expression. Conclutions : These results suggest that marine natural resources such as Porphyra thalli, Laminariae thallus, Ostreae concha, Sargassum thallus, Undaria thallus, Codium thalli, Syngnathoides biaculeatus and Hippocampus coronatus have anti-melanogenic effects, thereby exhibiting high potentials to be utilized as one of the ingredients for the development of new whitening functional cosmetics.