• Title/Summary/Keyword: tyrosinase protein expression

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Hypopigmenting Effects of Extracts from Bulbs of Lilium Oriental Hybrid 'Siberia' in Murine B16/F10 Melanoma Cells (백합뿌리 추출물의 멜라닌 생성 억제효과)

  • Yoon, Hoon Seok;Yang, Kyung-Wol;Kim, Jung Eun;Kim, Jeong Mi;Lee, Nam Ho;Hyun, Chang-Gu
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.5
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    • pp.705-711
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    • 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.

Whitening Effect of Androsace umbellata Extract (봄맞이 추출물의 미백 효능 연구)

  • Kim, Bo Yun;Park, Sung Ha;Park, Byoung Jun;Kim, Jin Jun
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.41 no.1
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    • pp.21-26
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    • 2015
  • The purpose of this study is to identify the whitening effect of Androsace umbellata extract. To discover the anti-pigmentation effective, we investigated Androsace umbellata extract on tyrosinase and melanogenesis inhibition. As results, it reduced tyrosinase activity and melanin contents in B16F1 melanoma cells in a dose-dependent manner with decreased to about 32% at a concentration of $25{\mu}g/mL$. To reveal how it works in inner-cellular level, we performed Western blot method. We found out that it also inhibited the protein expression in tyrosinase, tyrosinase related protein 1 (TRP-1), and microphthalmia associated transcription factor (MITF) in melanocytes. Therefore, we successfully identified the whitening effect of Androsace umbellata extract, and this finding suggested that Androsace umbellata extract is a considerable potent cosmetics ingredient for whitening. Based on this, we anticipated further researches about Androsace umbellata extract for gene levels and additional mechanisms to develop not only for functional cosmetics but for medicines or healthcare food.

Nypa fruticans wurmb Inhibits Melanogenesis via cAMP/PKA/CREB Signaling Pathway in B16 F10 Cells

  • So-Yeon Han;Hye-Jeong Park;Jeong-Yong Park;Seo-Hyun Yun;Mi-Ji Noh;Soo-Yeon Kim;Tae-Won Jang;Jae-Ho Park
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2021.04a
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    • pp.54-54
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    • 2021
  • The Melanoma Research Coalition reported melanoma affects humans of various races. This study was conducted to confirm the inhibitory effect of melanogenesis in B16 F10 cells of Nypa fruticans Wurmb of ethyl acetate fraction (NEF). Nypa fruticans Wurmb is an important component of the East Asian mangrove vegetation. It belongs to Araceae family. Traditionally, N. fruticans was used to treat various diseases such as asthma, sore throat, liver disease, a pain reliever, and can also be used as sedative and carminative. The present study, the inhibitory effect on melanogenesis was determined by Western blotting and RT-qPCR. The level of expression of tyrosinase, TRP-1, and TRP-2 is regulated by microphthalmia-associated transcription factor (MITF) and cAMP, and cAMP affects the activity of protein kinase A (PKA). Activated PKA stimulates the phosphorylation of cAMP-reactive element-binding protein (CREB) in the nucleus, thereby increasing the amount of MITF expression and enhancing melanogenesis. Western blotting and RT-qPCR analysis showed that NEF treatment decreased the expression of tyrosinase. Similarly, TRP-1 and TRP-2 levels were decreased, which were decreased significantly at compared with the untreated control. Also, NEF attenuated the IBMX mediated increase in the intracellular cAMP level and the phosphorylation of PKA. In conclusion, NEF significantly inhibited the expressions of melanogenesis through cAMP/PKA/CREB signaling pathways.

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Antioxidant Activities and Whitening Effects of a Mixture of the Eco-friendly Materials Pinus koraiensis and Hibiscus cannabinus L. (친환경 소재 잣나무 목재와 케나프 줄기 혼합물의 항산화 및 미백효과)

  • Oh, Min-Jeong;Yeom, Hyeon-Ji;Chae, Jung-Woo;Lee, Jin-Young
    • Journal of Life Science
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    • v.31 no.3
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    • pp.305-313
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    • 2021
  • This study verified the antioxidant and whitening activities of a Pinus koraiensis extract (PK) and a Hibiscus cannabinus L. extract (HC), and further evaluated the interaction of the extract ingredients when mixed at a 1:1 ratio (PKHC). The electron-donating and ABTS+ radical scavenging activities of the PKHC extract at 1,000 ㎍/ml concentration were 93.7% and 94%, respectively, indicating a higher efficacy than achieved with either extract alone. Measurements of the tyrosinase the activities in response to PK, HC, and PKHC extracts at 1,000 ㎍/ml concentrations showed inhibitions of 40%, 27.5%, and 43%, respectively, confirming a higher efficacy of the mixture due to the synergistic action of the ingredients. The cell toxicity values in melanoma cells treated with PK, HC, and PKHC at 1,000 ㎍/ml concentration were 87.4%, 80.2%, and 98%, confirming a higher viability in cells treated with the mixture due to antagonism. The expression of microphthalmia-associated transcription factor (MITF), tyrosinase-related protein-1 (TRP-1), tyrosinase-related protein-2 (TRP-2), and tyrosinase protein expression determined by Western blotting decreased by 53.9%, 64.8%, 67.3%, and 56.1%, respectively, when PKHC was administered at a concentration of 100 ㎍/ml. Reverse transcription-polymerase chain reaction (RT- PCR) results also showed that PKHC at a concentration of 100 ㎍/ml inhibited the mRNA expression of MITF, TRP-1, TRP-2, and tyrosinase mRNA by 54.4%, 64.9%, 66.6%, and 63.1%, respectively. Taken together, the data confirmed the antioxidant and whitening effect of the PKHC extract and verified the possibility that this extract mixture has great potential as a cosmetic ingredient.

Phenolic acids in Panax ginseng inhibit melanin production through bidirectional regulation of melanin synthase transcription via different signaling pathways

  • Jianzeng Liu ;Xiaohao Xu ;Jingyuan Zhou;Guang Sun ;Zhenzhuo Li;Lu Zhai ;Jing Wang ;Rui Ma ;Daqing Zhao;Rui Jiang ;Liwei Sun
    • Journal of Ginseng Research
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    • v.47 no.6
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    • pp.714-725
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    • 2023
  • Background: Our previous investigation indicated that the preparation of Panax ginseng Meyer (P. ginseng) inhibited melanogenesis. It comprised salicylic acid (SA), protocatechuic acid (PA), p-coumaric acid (p-CA), vanillic acid (VA), and caffeic acid (CA). In this investigation, the regulatory effects of P. ginseng phenolic acid monomers on melanin production were assessed. Methods: In vitro and in vivo impact of phenolic acid monomers were assessed. Results: SA, PA, p-CA and VA inhibited tyrosinase (TYR) to reduce melanin production, whereas CA had the opposite effects. SA, PA, p-CA and VA significantly downregulated the melanocortin 1 receptor (MC1R), cycle AMP (cAMP), protein kinase A (PKA), cycle AMP-response element-binding protein (CREB), microphthalmia-associated transcription factor (MITF) pathway, reducing mRNA and protein levels of TYR, tyrosinase-related protein 1 (TYRP1), and TYRP2. Moreover, CA treatment enhanced the cAMP, PKA, and CREB pathways to promote MITF mRNA level and phosphorylation. It also alleviated MITF protein level in α-MSH-stimulated B16F10 cells, comparable to untreated B16F10, increasing the expression of phosphorylation glycogen synthase kinase 3β (p-GSK3β), β-catenin, p-ERK/ERK, and p-p38/p38. Furthermore, the GSK3β inhibitor promoted p-GSK3β and p-MITF expression, as observed in CA-treated cells. Moreover, p38 and ERK inhibitors inhibited CA-stimulated p-p38/p38, p-ERK/ERK, and p-MITF increase, which had negative binding energies with MC1R, as depicted by molecular docking. Conclusion: P. ginseng roots' phenolic acid monomers can safely inhibit melanin production by bidirectionally regulating melanin synthase transcription. Furthermore, they reduced MITF expression via MC1R/cAMP/PKA signaling pathway and enhanced MITF post-translational modification via Wnt/mitogen-activated protein kinase signaling pathway.

New Whitening Agent From Pimpinella brachycarpa (참나물추출물의 멜라닌 생성저해 효과)

  • Kim, Jin-Hwa;Sim, Gwan-Sub;Lee, Dong-Hwan;Lee, Geun-Soo;Lee, Bum-Chun;Pyo, Hyeong-Bae
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.33 no.3
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    • pp.203-208
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    • 2007
  • To develop a new whitening agent for cosmetics from natural products, Pimpinella brachcarpa was selected for its inhibitory effect on melanogenesis in B16 melanoma cells. Crude ethanolic extract of P. brachycarpa and its four fractions-hexane, ethyl acetate(EtOAc), butanol and aqueous were evaluated for antioxidative effects and tyrosinase inhibitory activity. To elucidate the mechanism of active compounds of P. brachycarpa, we investigated the changes in protein level of tyrosinase, TRP-1 and TRP-2 using Western blotting and the changes in mRNA level of tyrosinase using RT-PCR technique. Following UV irradiation, expression of ET-1 in HaCaT keratinocytes was measured by quantitative enzyme immunoassay(EIA) using human ET-1 antibody. Crude ethanolic extract of P. brachycarpa and its four fractions-hexane, EtOAc, butanol and aqueous had free radical scavenging effect by 87.2, 2.5, 97.2, 80.5, 49.8% at 100 ${\mu}g/mL$ and tyrosinase inhibitory effect by 18.3, 15.1, 55.4, 13.1, 0 % at 100 ${\mu}g/mL$. P. brachycarpa EtOAc fraction significantly inhibited melanin production in B16 melanoma cells. Treatment with P. brachycarpa extract for 72 h suppressed the biosynthesis of melanin up to 58 % at 100 ${\mu}g/mL$. Especially, the EtOAc fraction of P. brachycarpa reduced the tyrosinase activity and tyrosinase expression in B16 melanoma cells in a dose-dependent manner. mRNA levels of tyrosinase and TRP-1 were markedly reduced by the EtOAc fraction of P. brachycarpa. Moreover, at the concentrations of $12.5{\sim}50{\mu}g/mL$ of the fraction, the production of UV-induced ET-1 in HaCaT keratinocytes(24 h after 8 $mJ/cm^2$ UVB irradiation) was reduced about 40%(p<0.05). P. brachycarpa could be used as a new natural skin-whitening agent due to the inhibitory effect of on melanin biosynthesis and endothelin-1 expression.

Inhibitory Effects of Resveratrol on Melanin Synthesis in Ultraviolet B-Induced Pigmentation in Guinea Pig Skin

  • Lee, Taek Hwan;Seo, Jae Ok;Baek, So-Hyeon;Kim, Sun Yeou
    • 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.

ERK Activation by Fucoidan Leads to Inhibition of Melanogenesis in Mel-Ab Cells

  • Song, Yu Seok;Balcos, Marie Carmel;Yun, Hye-Young;Baek, Kwang Jin;Kwon, Nyoun Soo;Kim, Myo-Kyoung;Kim, Dong-Seok
    • The Korean Journal of Physiology and Pharmacology
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    • v.19 no.1
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    • pp.29-34
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    • 2015
  • Fucoidan, a fucose-rich sulfated polysaccharide derived from brown seaweed in the class Phaeophyceae, has been widely studied for its possible health benefits. However, the potential of fucoidan as a possible treatment for hyperpigmentation is not fully understood. This study investigated the effects of fucoidan on melanogenesis and related signaling pathways using Mel-Ab cells. Fucoidan significantly decreased melanin content. While fucoidan treatment decreased tyrosinase activity, it did not do so directly. Western blot analysis indicated that fucoidan downregulated microphthalmia-associated transcription factor and reduced tyrosinase protein expression. Further investigation showed that fucoidan activated the extracellular signal-regulated kinase (ERK) pathway, suggesting a possible mechanism for the inhibition of melanin synthesis. Treatment with PD98059, a specific ERK inhibitor, resulted in the recovery of melanin production. Taken together, these findings suggest that fucoidan inhibits melanogenesis via ERK phosphorylation.

Inhibitory effects of Kirengeshoma koreana Nakai on Melanogenesis in B16F10 melanoma cells

  • Jang, Tae-Won;Choi, Ji-Soo;Mun, Jeong-Yun;Im, Jong-Yun;Nam, Su-Hwan;Kim, Do-Wan;Lee, Seung-Hyun;Park, Jae-Ho
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2019.04a
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    • pp.117-117
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    • 2019
  • Kirengeshoma koreana Nakai (K. koreana)was Saxifragaceae and rare plants in Korea, which is classified as an Critically Endangered (CR) species in Korea. Therefore, most of the studies on it were ecological and taxonomic, and there are no studies on biological activity. In this study, we evaluated the whitening activity of K. koreana extract (KKE). Melanogenesis Inhibitory effects were demonstrated by western-bot and RT-PCR for the effects of KKE on MITF, tyrosinase, TRP-1 and TRP-2 in IBMX-treated B16F10 melanoma cells. IBMX were reported as melanin synthesis enhancers. It could increase intracellular melanin synthesis by activation of the microphthalmia-associated transcription factor (MITF) signaling pathway. KKE showed no cytotoxicity at B16F10. In addition, KKE effectively inhibited the protein and mRNA levels of MITF, tyrosinase, TRP-1 and TRP-2. In conclusion, KKE inhibited melanin synthesis by inhibiting the expression of MITF and its downstream pathways tyrosinase, TRP-1 and TRP-2. Therefore, it was confirmed that K. koreana is a valuable resource for functional cosmetic and biomaterials.

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Melanogenesis Inhibitory Activity of Epicatechin-3-O-Gallate Isolated from Polygonum amphibium L.

  • Lee, Young Kyung;Hwang, Buyng Su;Hwang, Yong;Lee, Seung Young;Oh, Young Taek;Kim, Chul Hwan;Nam, Hyeon Ju;Jeong, Yong Tae
    • Microbiology and Biotechnology Letters
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    • v.49 no.1
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    • pp.24-31
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    • 2021
  • This study aimed to investigate the melanogenesis inhibitory activity of epicatechin-3-O-gallate (ECG) isolated from Polygonum amphibium L. ECG was isolated from the ethanol extract of P. amphibium L, and its chemical structure was determined using spectroscopic methods such as LC-ESI-MS, 1D-NMR, and UV spectroscopy. ECG inhibited the melanogenesis of B16F10 cells in a dose-dependent manner. Particularly, it decreased the melanin content by 27.4% at 200 µM concentration, compared with the control, in B16F10 cells, without causing cytotoxicity. It is noteworthy that the expression of three key proteins, including tyrosinase, tyrosinase-related protein-1 (TRP-1), TRP-2, and microphthalmia-associated transcription factor (MITF), involved in melanogenesis, is significantly inhibited by ECG. The ECG isolated in this study caused the inhibition of body pigmentation and tyrosinase activity in vivo in the zebrafish model. These results suggest that the ECG isolated from P. amphibium L. is an effective anti-melanogenesis agent.