• 동의생리병리학회지
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• 제21권6호
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• pp.1456-1461
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• 2007

Melanogenesis is induced mainly by ultraviolet radiation of sunlight and ${\alpha}-melanocyte$-stimulating hormone (${\alpha}-MSH$) which binds to a specific G protein coupled receptor. The purpose of this study was to investigate the mechanism of melanogenesis inhibition in B16/F10 cells by methanol extract of Rubus coreanus Miquel (RCM). In the present study, ${\alpha}-MSH$ and forskolin led to a stimulation of melanin synthesis that appeared to result from an increased tyrosinase activity and melanin content. However, RCM inhibited the ${\alpha}-MSH$- and forskolin-induced melanin synthesis. In addition, RCM abolished the ${\alpha}-MSH$- and forskolin-induced cytoplasmic dendricity. Regarding protein levels of the melanogenic enzymes, the amounts of tyrosinase and tyrosinase-related protein 1 (TRP-1) were increased after incubation with α-MSH and forskolin. The treatment of RCM decreased the ${\alpha}-MSH$- and forskolin-induced expression levels of tyrosinase and TRP-1. Based on these findings, it is likely that RCM exerts its depigmenting effects in B16/F10 cells through the suppression of tyrosinase and TRP-1 expression, which are key enzymes for melanogenesis.

• Journal of Periodontal and Implant Science
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• 제31권2호
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• pp.357-369
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• 2001

In this present study, the healing process and the recurrence of pigmentation were evaluated clinically and histologically in accordance with the extent and the range of pigmentation after phenol was applied to remove melanin pigmentation in gingiva. Six mongrel dogs were used. The melanin pigmentation in canine gingiva were classified into slight, moderate and severe according to the extent of pigmentation and divided into local and diffuse types according to the range. Following general and local anesthesia, 90% phenol was applied to the pigmented gingiva of the subjects with small cotton balls until the surface was etched to be whitish and was neutralized with small cotton balls soaked by 95% alcohol. The contralateral pigmented gingiva to the one treated with phenol, was treated by surgical deepithelialization. At 1, 3 and 8 weeks, the treated gingiva was examined clinically and evaluated histologically following H-E stain, and HMB 45 stain for melanocyte after biopsy. In the phenol treated sites, epithelium and connective tissue healed normally and there was no pigmentation at 1 week. At 3 weeks of healing, melanin repigmentation was observed in the severe local type and moderate to severe diffuse type. In the surgically deepithelialized sites, healing was delayed, compared to phenol treated sites and the infiltration of the inflammatory cells and congestion in connective tissue was shown at 1 week. At 3 weeks, healing was completed and there was a partial melanin repigmentation. At 8 weeks of healing, the extent and the range of repigmentation were increased in both group according to the extent or range priot to depigmentation procedure. These results suggpriorest that the removal of melanin pigmentation with 90% phenol application result in normal healing process of gingiva. However, in the severe local type and moderate to severe diffuse type, sites treated with phenol showed repigmentation at 3 week, which was earlier than surgical deepithelialized sites. Therefore it is required to select appropriate method according to initial condition of pigmentation.

• Applied Microscopy
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• 제28권2호
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• pp.127-137
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• 1998

This study was observed of the skin that changed after irradiation of the ultraviolet A. All the mouse were hairless which the weight are about 25g and the ages $6\sim8$ weeks old. The mouse were divided into six groups; control, irradiated for 6 hours, 3 days, 7 days, 14 days, 21 days and 28 days. Each group was irradiated with ultraviolet that is $320nm\sim366nm$ of wavelengths. After irradiated, the skin was observed with the electron microscope and the light microscope. The results are as follow: 1) Light microscopy With following irradiation, the epidermis was not changed to most groups but at the 28 days group was thickened and deposit the melanocyte. The elastic fibers within the epidermis were thickened and twisted with following irradiation. 2) Eelectron microscopy The elastic fibers were slightly clumped at 6 hours group, mildly increased and partly aggregated in the 3 days group, branched and tangled at 7 days group, irregulated and electron density at 14 days group, sightly thickened and twisted at 21 days group, and randomly arranged, shortened, twisted, and electron density at 28 days group.

• 생약학회지
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• 제45권3호
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• pp.262-267
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• 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.

• Food Science and Biotechnology
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• 제17권4호
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• pp.700-704
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• 2008

Anti-obesity effects of $\beta$-cyclodextrin in obese C57BL/6J mice induced by a high fat diet (HD) were observed. The administration of $\beta$-cyclodextrin reduced the gain of body weight, abdominal fat, liver weight, the lipid deposits of hepatocytes and the size of adipocytes in the HD group. In serum analysis, the total and low-density lipoprotein-cholesterols were significantly decreased in the $\beta$-cyclodextrin-supplemented HD group than in the HD group. However, high-density lipoprotein-cholesterol was not changed in these groups. In hypothalamic homogenates, the decrease of neuropeptide Y and increase of $\alpha$-melanocyte stimulating hormone were detected in the $\beta$-cyclodextrin-supplemented HD group compared to that in the HD group. These effects of $\beta$-cyclodextrin were similar to those of Garcinia cambogia, which is widely used as a natural anti-obesity product. These results suggest that $\beta$-cyclodextrin has anti-obesity effects through the lowering of the abdominal fat pad and inhibits the central effects of hunger.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2019년도 추계학술대회
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• pp.90-90
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• 2019

Hibiscus syriacus exhibited promising potential as a new source of food and colorants containing various anthocyanins. However, the function of anthocyanins from H. syriacus has not been investigated. In the current study, we evaluated whether anthocyanins from the H. syriacus varieties Pulsae and Paektanshim (PS and PTS) inhibit melanin biogenesis. B16F10 cells and zebrafish larvae were exposed to PS and PTS in the presence or absence of ${\alpha}$-melanocyte-stimulating hormone (${\alpha}$-MSH), and melanin contents accompanied by its regulating genes and proteins were analyzed. PS and PTS moderately downregulated mushroom tyrosinase activity in vitro, but significantly decreased extracellular and intracellular melanin production in B16F10 cells, and inhibited ${\alpha}$-MSH-induced expression of microphthalmia-associated transcription factor (MITF) and tyrosinase. PS and PTS also attenuated pigmentation in ${\alpha}$-MSH-stimulated zebrafish larvae. Furthermore, PS and PTS activated the phosphorylation of extracellular signal-regulated kinase (ERK), whereas PD98059, a specific ERK inhibitor, completely reversed PS- and PTS-mediated anti-melanogenic activity in B16F10 cells and zebrafish larvae, which indicates that PS- and PTS-mediated anti-melanogenic activity is due to ERK activation. Moreover, chromatography data showed that PS and PTS possessed 17 identical anthocyanins as a negative regulator of ERK. These findings suggested that anthocyanins from PS and PTS inhibited melanogenesis in vitro and in vivo by activating the ERK signaling pathway.

• 대한화장품학회지
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• 제28권1호
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• pp.135-149
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• 2002

To date, research on the regulation of melanogenesis has focused on factors which affect tyrosinase, the rate-limiting enzyme in the melanogenic pathway, by searching for chemicals which competitively inhibit tyrosinase function. Many types of tyrosinase inhibitors have been developed, but no satisfactory results have been made clinically until now, To find a new whitening agent, which effectively inhibits melanogenesis, we synthesized several compounds and selected compounds by cell-based assay system. Finally, 3, 4, 5-trimethoxy cinnamaie thymol ester(TCTE, Melasolv) was selected and the effects of TCTE on melanogenesis were investigated. Treatment of mouse-derived melanocyte melan-a cells with TCTE results in a marked down-regulation of tyrosinase activity. 80% decrease of tyrosinase activity occurs with 30uM TCTE treatment for 72 hours without affecting cell growth. The inhibition of tyrosinase activity is dose-dependent and melanin content was also decreased to 40%. From the in vitro tyrosinase assay using cell extract, TCTE does not act as a direct inhibitor of the enzyme. Treatment of melan-a cultures with TCTE blocks the increase in tyrosinase activity by either forskolin, 3-isobutyl-1-methtyl-xanthine. TCTE decreased the expression of tyrosinase, TRP-1 without effects on TRP-2 protein expression through the down regulation of tyrosinase and TRP-1 mRNA. From the results of cAMP immunoassays, intracellular levels of the cyclin nucleotide are unaffected in cells treated with TCTE. The inhibitory effects of melanin synthesis were also shown in reconstitute human epidermis model by topical application. These findings suggest that TCTE can be used for studying the regulation of melanogenesis and depigmenting agent.

• 대한화장품학회지
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• 제34권3호
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• pp.183-188
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• 2008

미백제를 선발하기 위해 주로 사용하는 현재의 방법은 in vitro 타이로시네이즈 활성 및 항산화능을 측정하는 것이다. 이 결과에 기초하여 다음 단계인 멜라노사이트에서의 멜라닌 생성량을 측정한다. 세포 내의 멜라닌 생성량 측정 법은 시간, 인력 및 숙련도가 요구된다. 따라서 초기 선발 방법의 신뢰성이 중요하다. 200개 중국시료 중 측정범위 내에서 세포독성이 없는 34개를 대상으로 세포 내 멜라닌량, 타이로시네이즈 활성, 항산화능의 상관관계를 조사하였다. 조사결과 직선의 상판관계를 확인할 수 없었다. 이 결과는 현재 선발방법의 한계 및 새로운 방법이 필요함을 보여주었다.

• 한국식품저장유통학회지
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• 제18권6호
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• pp.998-1001
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• 2011

신선초 추출물의 미백활성을 확인하고 이의 소재를 개발하기 위하여, tyrosinase 저해 활성 및 B16F10 세포주를 이용한 melanogenesis 저해능을 확인하였다. 그 결과, 신선초 50% 에탄올 추출물에서 B16F10 melanoma 세포의 멜라닌 생성을 억제하였다. Melanogenesis 억제 활성에 관련이 있는 분자마커의 확인을 위하여 RT-PCR에 의한 mRNA 발현 수준을 검토한 결과, 농도의존적으로 TYR, TYRP-1. TYRP-2의 발현이 억제되었으며 MITF 전사인자의 발현도 감소시켰다. 본 추출물은 독성이 없으므로, 향후 신선초 추출물에서의 미백활성을 나타내는 단일물질을 분리/조제에 응용한다면 cosmeceutical의 소재로 개발이 기대된다.

• 약학회지
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• 제47권1호
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• pp.31-36
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• 2003

To investigate the effect of protein kinase on melanin production via cAMP-dependent pathway, we measured the melanin amount and tyrosinase activity in B16 melanoma cells stimulated by alpha-melanocyte stimulating hormone (MSH), forskolin and 8-Br-cAMP. MSH, forskolin and 8-Br-cAMP significantly increased both melanin production and tyrosinase activity in B16 cells. Melanin production and tyrosinase activity by MSH are significantly inhibited by cyclic AMP-dependent protein kinase inhibitor (KT5720) and protein kinase C down-regulation treated with PMA. Bisindolmaleimide (1$\mu$M), protein kinase C inhibitor, significantly inhibited melanin production and tyrosinase activity stimulated by MSH, forskolin and 8-Br-cAMP with the following order of potency: MSH＞forskolin＞8-Br-cAMP. Tyrosine kinase inhibitor, genistein and DHC, significantly inhibited both, but the inhibitory effect was more potent in 8-Br-cAMP-stimulated B16 cells than MSH-stimulated cells. NFkB inhibitor (parthenolide) significantly inhibited melanin production and tyrosinase activity. Neither melanin production nor tyrosinase activity induced by MSH, forskolin and 8-Br-cAMP were affected by KN-62 (calmodulin-dependent protein kinase II inhibitor), PD098059 (mitogen-activated protein kinase inhibitor, MAPKK) and worthmannin (phosphatidylinositol 3-kinase inhibitor). These results suggest that both protein kinase C and tyrosine kinase are involved in melanin production by cyclic AMP-dependent pathway and NFkB pathway may play an important role in cyclic AMP-dependent melanin production in B16 melanoma cells.