• 동의생리병리학회지
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• 제30권1호
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• pp.47-53
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• 2016

Kaempferia galanga L. is one of the plants in Zingiberaceae family. It is used by people in many regions of Asia and Africa for relieving toothache, abdominal pain, muscular swelling and rheumatism. Tyrosinase is a key enzyme for melanogenesis, and hyperpigmentation is associated with abnomal accumulation of melanin pigment. This study aimed to investigate the inhibition of melanogenesis by hexane extract of Kaempferia galanga L. (HKG) in B16F10 melanoma cells. Cell-free tyrosinase, melanin contents, intracellular tyrosinase activity and western blot analysis were performed to elucidate the effects on anti-melanogenesis. Cytotoxicity of the extracts was measured using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and determined the concentration of 12.5, 25 μg/ml. HKG significantly inhibited to activities of intracellular tyrosinase and melanin synthesis in the absence or presence of α-melanocyte stimulating hormone (α-MSH) with dose-dependent manner. And HKG inhibited the expression of tyrosinase, tyrosinase-related protein 1 (TRP-1) and tyrosinase-related protein 2 (TRP-2), regardless of the presence or absence of α-MSH. HKG also down-regulated phosphorylation of p38 and JNK, and up-regulated phosphorylation of Akt. These effects were not related to its cytotoxicity action. These results indicate that HKG has the potential to be a useful therapeutic agent for treating hyperpigmentation disorders and as a beneficial additive in whitening agents in cosmetics industry.

• Preventive Nutrition and Food Science
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• 제21권2호
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• pp.155-159
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• 2016

Previous research showed that resveratrol (trans-3,4',5-trihydroxystilbene) and pinostilbene (trans-3-methoxy-4',5-dihydroxystilbene) were able to inhibit tyrosinase directly; however, anti-melanogenic effects of pterostilbene (trans-3,5-dimethoxy-4'-hydroxystilbene) and resveratrol trimethyl ether (RTE) have not been compared. To investigate the hypopigmentation effects of pterostilbene and RTE, melanin contents and intracellular tyrosinase activity were determined by western blot analysis. Firstly, pterostilbene showed the inhibitory effects on ${\alpha}$-melanocyte stimulating hormone (MSH)-induced melanin synthesis stronger than RTE, resveratrol, and arbutin. Pterostilbene inhibited melanin biosynthesis in a dose-dependent manner in ${\alpha}$-MSH-stimulated B16/F10 murine melanoma cells. Specifically, melanin content and intracellular tyrosinase activity were inhibited by 63% and 58%, respectively, in response to treatment with $10{\mu}m$ of pterostilbene. The results of western blot analysis indicated that pterostilbene induced downregulation of tyrosinase protein expression and suppression of ${\alpha}$-MSH-stimulated melan-A protein expression stronger than RTE or resveratrol. Based on these results, our study suggests that pterostilbene can induce hypopigmentation effects more effectively than resveratrol and RTE, and it functions via downregulation of protein expression associated with hyperpigmentation in ${\alpha}$-MSH-triggered B16/F10 murine melanoma cells.

• 한방안이비인후피부과학회지
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• 제16권3호
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• pp.129-144
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• 2003

Recently many efforts were focused to understand the mechanical insights of melanogenesis to develop the agents for hyper-pigmentation and hypo-pigmentation. In the melanin biosynthetic pathway, tyrosinase is the rate limiting enzyme, and ${\alpha}$-melanocyte stimulating hormone(MSH) or cAMP-elevating agents stimulate melanogenesis and enhance the melanin synthesis and the tyrosinase activity. The author has analyzed the effects of Rhizoma Bletillae on the basal melanogenic activities of B16／F10 mouse melanoma cells, and on the ${\alpha}$-MSH or forskolin-induced melanogenesis. Rhizoma Bletillae alone markedly suppressed melanin content and tyrosinase activity in a dose-dependent manner. Pretreatment of the cells with Rhizoma Bletillae also suppressed the increase of ${\alpha}$-MSH (100 nM) or forskolin (20 ${\mu}M$)-induced melanin content and tyrosinase activity. The decrease in the tyrosinase activity was paralled by a decrease in the abundance of tyrosinase protein and tyrosinase promoter activity. Pretreatment of the cells with Rhizoma Bletillae also inhibited the increase of forskolin(20${\mu}M$) induced the amount of tyrosinase protein and tyrosinase promoter activity. The results of DOPA staining revealed that pretreatment of the cells with Rhizoma Bletillae showed less intensity than B16 melanoma cells stimulated with ${\alpha}$-MSH or forskolin. These results suggest that Rhizoma Bletillae inhibits melanogenesis and abrogates ${\alpha}$-MSH and cAMP-induced melanogenesis in B16 melanoma cells.

• Bulletin of the Korean Chemical Society
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• 제34권10호
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• pp.3017-3021
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• 2013

Coumaroyl dipeptide amide, Coumaric acid-LG-$NH_2$, was prepared successfully using the solid-phase method, and its efficacy as a skin whitening agent was studied. Coumaric acid-LG-$NH_2$ was prepared with Rink-amide resin, and 96.354% of purity was obtained. Using MTT assay and LDH release assay, we found that it exhibited very low cytotoxicity. And, we found that Coumaric acid-LG-$NH_2$ inhibited tyrosinase activity dose-dependently and showed superior tyrosinase inhibitory activity to well-known whitening agent, arbutin. $IC_{50}$ value of Coumaric acid-LG-$NH_2$ was 182.4 ${\mu}M$, and $IC_{50}$ value of arbutin was 384.6 ${\mu}M$. Also, in measurement of melanin contents using B16F1 melanoma cell lines, Coumaric acid-LG-$NH_2$ reduced melanin production induced by ${\alpha}$-MSH statistically significant, and showed superior melanin inhibitory activity to p-coumaric acid or arbutin. In addition, Coumaric acid-LG-$NH_2$ reduced MC1R mRNA expression level. Thus, we concluded that MC1R pathway is the significant pathway of Coumaric acid-LG-$NH_2$, and Coumaric acid-LG-$NH_2$ has great potential to be used as novel whitening agents.

• 동의생리병리학회지
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• 제20권5호
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• pp.1271-1274
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• 2006

We investigated the effects of aqueous extracts from twelve medical herbs on ${\alpha}$-melanocyte stimulating hormone (${\alpha}$-MSH)-induced melanogenesis in B16F10 mouse melanoma cell. The cells were incubated with ${\alpha}$-MSH and aqueous extracts 5 days and 2 days and then analysed melanin amount and tyrosinase activities, respectively. Nine aqueous extract of herbs examined at 1 mg/m${\ell}$ level decreased melanin synthesis in B16F10 cell, especially in Agastache rugosa, Leonurus siviricus and Murus bombycis. The significant decrease of released extracellular melanin were also observed in treated cells with aqueous extract from Leonurus siviricus, Murus bombycis and Ledebouriella seseioides. The ${\alpha}$-MSH-induced activation of tyrosinase was inhibited in cells treated with aqueous extract from Cuscutae semen, Angelica tenuissima and Agastache rugosa. These results suggest that herbs inhibiting melanogenesis through tyrosinase activity may apply to develop whitening drugs and cosmetics.

• 대한화장품학회지
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• 제41권4호
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• pp.383-389
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• 2015

본 연구는 은행 열매 오일의 멜라닌 생성 억제 효과를 확인한 것이다. 은행나무 열매 오일은 DPPH assay와 FRAP assay를 사용하여 라디컬 소거능을 시험하였다. 결과적으로 은행나무 열매 오일은 DMSO를 용매로 0.06% 녹였을 때, DPPH assay에서 9.96% 소거활성을 나타내었고 FRAP는 1.33 mM의 ferric sulfate ($FeSO_4$)를 생성하였다. 은행 열매 오일은 tyrosinase inhibition assay에서 37.72%의 억제력을 가졌고 B16/F10 세포에 멜라닌 생합성 실험을 통해 확인하였다. 은행 오일 0.06%에서 ${\alpha}$-MSH 처리 구에 비해 48.02%의 멜라닌 생성을 억제하였다. Tyrosinase, tyrosinase related protein-1 (TRP-1), tyrosinase related protein-2 (TRP-2), microphthalmia-associated transcription factor (MITF)의 유전자 발현 수준은 control군에 비해 0.04%와 0.06% 농도 군이 크게 감소하였다. 결과적으로 은행 열매 오일 추출물이 멜라닌 생성을 억제하는 효과가 있는 것으로 확인되었다.

• 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.

• Journal of Applied Biological Chemistry
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• 제54권4호
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• pp.284-289
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• 2011

선학초 추출물을 기능성 화장품 소재로 활용하기 위하여 미백효과를 검증하였다. 세포 내 tyrosinase 저해활성 측정결과 선학초 에탄올 추출물 $500{\mu}g/mL$에서 42%의 저해활성을 나타내었다. 이는 선학초 추출물이 세포 내 tyrosinase 발현을 억제시킴으로서 멜라닌 합성 또한 저해 하는 것임을 확인 할 수 있었다. 선학초 추출물의 단백질 발현과 mRNA 발현 억제효과를 검토한 결과 선학초 열수 및 에탄올 추출물을 처리한 B16F10군에서는 tyrosinase protein의 발현이 처리하지 않은 군보다 감소함을 확인할 수 있었다. 결과적으로 선학초 추출물의 미백효능을 확인할 수 있었으며, 식품 및 화장품의 기능성 소재로 이용이 가능할 것으로 판단된다.

• 한방안이비인후피부과학회지
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• 제23권3호
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• pp.11-32
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• 2010

Objective : The aim of this study is to determine the effects of Dendrobii herba extract and Punica granatum extract on skin disease and skin beauty. Methods : To investigate in vitro anti-oxidant activity assay, ethanol extracts of medicinal plants tested by DPPH radical, xanthine oxidase activity. In the next experiment, to investigate anti-inflammatory activity assay, examined by relations in NO synthesis, IL-$1{\beta}$, IL-6, TNF-${\alpha}$, NF-${\kappa}B$, COX-2, MAP kinase. To study Skin wrinkle formation effect, we were examined by tyrosinase activities, melanin synthesis in MNT-1 cell. Results : 1. In an anti-oxidant test, Dendrobii and Punica granatum extract showed high radical scavenging activity. 2. In an anti-inflammatory test, Dendrobii herba and Punica granatum extract weakly inhibited the lipopolysaccharide(LPS)-induced nitric oxide(NO) release from RAW 246.7 macrophage cells. Dendrobii herba and Punica granatum extract also inhibited LPS-induced IL-$1{\beta}$ and COX-2 expressions. The inhibitory effect of Dendrobii herba and Punica granatum extract on macrophage activation were via the inhibition of NF-${\kappa}B$, evidenced by transient transfection assay. however, Dendrobii herba and Punica granatum extract did not have any effects about activation of Jun-N-terminal kinase(JNK) and inhibition of p38 MAP kinase in RAW 264.7 cells. 3. In the skin wrinkle formation assay, Dendrobii herba and Punica granatum extract weakly inhibited collagenase and elastase, however it was not statistically significant. 4. In the skin whitening assay, Dendrobii herba and Punica granatum extract weakly inhibited tyrosinase activity, however, it was not statistically significant. They did not have any effect on melanin synthesis, indicating that they could not be applicable for skin whitening. Conclusion : Dendrobii herba extract and Punica granatum extract may play a significant role in skin disease and skin beauty.

• 대한화장품학회지
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• 제32권4호
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• pp.219-225
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• 2006

본 연구는 항유 추출물의 화장품 원료로서의 특성을 알아보기 위하여 항산화, 미백 및 항염 효과와 관련된 다양한 실험을 실시하였다. 30%, 70% 그리고 100% 메탄올로 추출한 각각의 향유 추출물을 대상으로 실험한 결과 DPPH 라디칼 소거 효과는 30%와 70% 추출물들은 0.025% 이상의 농도에서, 100% 추출물은 0.1% 이상의 농도에서 80%의 효과를 나타내었다. 세포내 멜라닌 생성 억제 효과는 각각의 용매별 추출물 모두 0.1% 이상의 농도에서 80%의 효과를 나타내었다. 이후 70% 메탄을 추출물을 대상으로 MPLC를 사용하여 성분 분리 실험을 실시한 결과 4개의 분획들을 얻었고, 이들을 대상으로 실험한 결과 1번, 2번 그리고 3번 분획들에서 항산화(DPPH 라니칼 소거, Mn-SOD 생성 억제), 미백(멜라닌 생성 억제) 그리고 항염($IL-1{\alpha}$, IL-6, COX-2, Total NO 생성 억제)효과를 나타내었다.