• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
• /
• v.24 no.1
• /
• pp.25-35
• /
• 2011

Objective : The present study was designed to assess the potential inhibitory activity of an ethanol extract of Belamcandae Rhizoma (EBR) on the alpha-melanocyte stimulating hormone (${\alpha}$-MSH)-induced melanogenesis signal pathway in B16F10 melanoma cells. Methods : Several experiments were performed in B16F10 melanoma cells. We studied tyrosinase activity, melanin content, cell-free tyrosinase activity and DOPA stain, and performed Western blots and RT-PCR for proteins and mRNA involved in melanogenesis. Results : ${\alpha}$-MSH-induced tyrosinase activity and melanin content were inhibited significantly by EBR. EBR markedly suppressed the protein expression level of tyrosinase in B16F10 melanoma cells. On the other hand, the expression of tyrosinase-related protein-1 (TRP-1) and -2 (TRP-2; DCT) were not affected by EBR. To elucidate the mechanism of the depigmenting property of EBR, we examined the involvement EBR in cAMP response element binding (CREB) protein phosphorylation and microphthalmia-associated transcription factor (MITF) signalling induced by ${\alpha}$-MSH. EBR did not regulate CREB phosphorylation and MITF expression by ${\alpha}$-MSH. Nevertheless, the mRNA expression of tyrosinase was significantly attenuated by EBR treatment without changes in the expression of TRP-1 and -2 mRNA. Conclusion : Our study suggested that EBR inhibits ${\alpha}$-MSH-induced melanogenesis by suppressing tyrosinase mRNA.

• Proceedings of the SCSK Conference
• /
• 2003.09a
• /
• pp.611-616
• /
• 2003

Korean ginseng(Panax Ginseng C.A. Meyer) known as a oriental miracle drug is an important medicinal plant. Ginseng has been used for geriatric, tonic, stomachic, and aphrodisiac treatments for thousands years. Also, it is an antibiotic and has therapeutic properties against stress and cancer. Ginseng is widely distributed all over the world. Among them, Korean mountain ginseng has the most valuable effect on pharmaceuticals. The roots of mountain ginseng contained several kinds of ginsenosides that have many active functions for the human body. However, the study of mountain ginseng has a limit because the mountain ginseng is very expensive and rare. So, we artificially cultured mountain ginseng adventitious roots using the bioreactor culture system. We induced callus from original mountain ginseng, directly dug up in mountain and aged about one hundred ten years. Separated adventitious roots were precultured in 500ml conical flasks and then, transferred in 20L bioreactors. The adventitious roots of mountain ginseng were harvested after culturing for 40days, dried and then, extracted with several solvents. In this study, we investigated the whitening effect, anti-wrinkle effect and the safety of tissue cultured adventitious roots extract of mountain ginseng in order to identify the merit as a cosmetic ingredient. Particularly, extract of mountain ginseng adventitious roots showed whitening and anti-wrinkle effects. The inhibitory effect of this extract on the melanogenesis was examined using B-16 melanoma cell. When B-16 melanoma cells were cultured with adventitious root extract, there was a dramatically decrease in melanin contents of 8-16 melanoma cell. And we identified this extract inhibited Dopa auto-oxidation significantly. Also, when transformed mouse fibroblast L929 cells were treated with this extract, there was a significant increase in collagen synthesis. The results show significant inhibited melanization and wrinkle without inhibiting cell viability.

• Journal of environmental and Sanitary engineering
• /
• v.22 no.1 s.63
• /
• pp.75-83
• /
• 2007

Clove extract by methanol increased expression of the tyrosinase gene on B16 mouse melanoma cells containing tyrosinase promoter. $10{\mu}g/mL$ and $100{\mu}g/mL$ of the extract showed expression rate of the tyrosinase gene about 138% and 245%, respectively, compared with control. At $500{\mu}g/mL$, expression rate of the extract was impossible to measurement by high cytotoxicity. The solvent fraction of methylene chloride also exhibited highly expression rate as methanol extract. However, the solvent fractions of butyl alcohol and water showed repressive effect on expression of tyrosinase gene at $500{\mu}g/mL$. In MTT assay, cell survival rate of the extract exhibited similar to expression rate of tyrosinase gene. That is, $10{\mu}g/mL$ and $100{\mu}g/mL$ of the extract showed the cell survival rate about 128% and 187%, respectively.

• Journal of Life Science
• /
• v.27 no.3
• /
• pp.318-324
• /
• 2017

This study was performed to improve the antioxidant and skin-whitening activities of 70% ethanol extract from Sesamum indicum L. (SIL). The electron-donating ability of the SIL extract was 71.7% at a concentration of $1,000{\mu}g/ml$. The whitening effects that was measured by tyrosinase inhibition assay. As a result, SIL extract was shown 42% at $1,000{\mu}g/ml$ concentration. The cell toxicity on B16F10 melanoma cells of SIL of 70% ethanol extract showed 84.3% at $1,000{\mu}g/ml$ concentration. The microphthalmia-associated transcription factor (MITF), tyrosinase relate protein-1 (TRP-1), tyrosinase relate protein-2 (TRP-2) and Tyrosinase protein and mRNA expression inhibitory effect of SIL extract were measured by western blot and reverse transcription- polymerase chain reaction (PCR) at 50, 250, $500{\mu}g/ml$ concentration. Consequently, the MITF, TRP-1, TRP-2, Tyrosinase protein expression inhibitory effect of SIL extract was decreased by 68.3%, 39.2%, 89.7%, 22.3%, respectively, at $500{\mu}g/ml$ concentration. Moreover, MITF, TRP-1, TRP-2, Tyrosinase mRNA expression inhibitory effect by reverse-transcription-PCR of SIL extract was decreased by 81.8%, 66.5%, 84.2%, 68.1%, respectively, at $500{\mu}g/ml$ concentration. Therefore, we excellently identified the antioxidant activities and whitening effect of SIL extract, and this finding suggested that SIL extract has great potential as a cosmetic ingredients.

• The Korea Journal of Herbology
• /
• v.29 no.6
• /
• pp.157-163
• /
• 2014

Objectives : Gamisoyo-san complex prescription were made with Angelicae Gigantis Radix, Paeoniae Radix, Atractylodes rhizome white, Hoelen, Bupleuri Radix, Moutan Cortex Radicis, Gardeniae Fructus, Zingiberis Rhizoma Crudus, Menthae Herba. The purpose of this study was to research the whitening effect of the extract from Gamisoyo-san, which is one of the used herbal complex prescription. Methods : This study investigated inhibitory effect of Gamisoyo-san in tyrosinase activity. Cell viability were performed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Then, Gamisoyo-san measured reversed-transcription-PCR for mRNA expression using B16F10 mouse melanoma cells. Results : For whitening effects, the tyrosinase inhibition effect of extract was shown to 52.4% at $5,000{\mu}g/m{\ell}$ concentration. The cell viability on B16F10 melanoma cells of Gamisoyo-san extract showed higher than 75% at $1,000{\mu}g/m{\ell}$ concentration. In this study, an experiment was performed by setting the non-toxic concentration range of 50, 150, $250{\mu}g/m{\ell}$. The Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as a positive control. The microphthalmia-associated transcription factor (MITF), tyrosinase related protein-1 (TRP-1), tyrosinase related protein-2 (TRP-2), tyrosinase mRNA expression inhibitory by reverse transcription-PCR of Gamisoyo-san extract were decreased by 95.3%, 98.8%, 96.3% and 49.5% at $250{\mu}g/m{\ell}$ which the highest concentration. Conclusions : All these findings could verify that whitening effects of Gamisoyo-san extract by tyrosinase inhibitory activity and mRNA expression. The Gamisoyo-san could be used as material for functional cosmetics, such as skin whitening products.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.34 no.9
• /
• pp.1325-1329
• /
• 2005

To estimate the inhibitory effect of Lithospemum erythrorhizon root extract on melanin biosynthesis, we tested its inhibitory effects on tyrosinase promoter in B16 mouse melanoma cells. Lithospermum erthrorhizon root extract had inhibitory effect above $33\%$ on tyrosinase promoter at $10{\mu}g/mL$ and exhibited no cytotoxicity under $100{\mu}g/mL$. Also, melanin biosynthesis decreased approximately $11\%$ and $24\%$ at $10{\mu}g/mL$ and $100{\mu}g/mL$, respectively. Therefore, Lithospermum erythrorhizon root extract would be considered very effective regulator of tyrosinase promoter and melanin biosynthesis.

• KSBB Journal
• /
• v.25 no.5
• /
• pp.478-482
• /
• 2010

Inhibitory melanogenesis by Bambusae caulis in Teaniam (Phyllostaachys nigra var. henonis Stapf) was studied. Tyrosinase inhibition activities were evaluated with six different extracts. Among them the extract with methanol showed the highest tyrosinase activity inhibition. MTT assay with B16 melanoma showed that the extract was not toxic up to the concentration of 50 ppm. The melanogenesis was clearly inhibited by the extract when it was examined by the melanin content assay in the cell. When the extract was dosed as 10 ppm, the melanogenesis was reduced to 68% in culture medium and 74% in the cell. By the proteome analysis with 2-D electrophoresis, 171 protein spots were found in the control gel and 282 spots were detected in the sample gel. Among 120 spot proteins matched, 12 spots were identified as proteins involved in the melanogenesis mechanism.

• Journal of Acupuncture Research
• /
• v.21 no.6
• /
• pp.63-84
• /
• 2004

Objective & Methods : To study the effects of anti-cancer, anti-metastasis and immune response improvement of herbal-acupuncture with Eclipta prostrata diffusae herba infusion solution(ELP-HAS), we injected ELP-HAS into Chung-wan(CV12) of C57BL/6 mice implanted intravenously with B16-F10 melanoma. We have reached the following conclusion through the effect on the percentage of $CD25^+/CD4^+$, $CD8^+/CD3e^+$, $CD69^+/B220^+$, $NK1.1^+/CD3e^+$cells in mouse PBMCs, the effect on the pulmonary colony formation number, and the effect on MST(Median Survival Time) and ILS(Increase of Life Span) of C57BL/6 mice implanted intravenously with B16-F10 melanoma. Results : The results were obtained as follows : 1. In the experiment groups treated with ELP(Ecliptae Herba) Herbal acupuncture, the spleen cell proliferation in BALB/c mouse was significantly increased compared with control group. 2. In the experiment groups treated with ELP(Ecliptae Herba) Herbal acupuncture, the percentage of $CD25^+/CD4^+$, $CD8^+/CD3e^+$, $CD69^+/B220^+$, $NK1.1^+/CD3e^+$cells in C57BL/6 mouse PBMCs was increased compared with control group. 3. In the experiment groups treated with ELP(Ecliptae Herba) Herbal acupuncture, the pulmonary colony formation number of C57BL/6 mice implanted intravenously with B16-F10 melanoma was decreased significantly compared with control group. 4. In the experiment groups treated with ELP(Ecliptae Herba) Herbal acupuncture, MST(Median Survival Time) and ILS(Increase of Life Span) of C57BL/6 mice implanted intravenously with B16-F10 melanoma were increased significantly compared with control group. Conclusions : ELP Herbal acupuncture injection into Chung-wan(CV12) of C57BL/6 mice is considered to be effective in anti-cancer, anti-metastasis and immune response improvement.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.20 no.5
• /
• pp.1271-1274
• /
• 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.

• Journal of Ginseng Research
• /
• v.18 no.3
• /
• pp.151-159
• /
• 1994

To evaluate the anticarcinogenic effect and its mechanism of red ginseng, the mice were treated with red ginseng and received subcutaneous Bl6 melanoma cell line injection on the back. Tumor incidence was same (100%) both in water and red ginseng-treated groups, but tumor production was delayed in red ginseng-treated group. Survival time was somewhat longer in red ginseng-treated group. The histopathological findings were similar in both groups, but lymphocytic infiltration around the tumor and melanin production in the tumor cells were prominent in the red ginseng-treated group. Flow cytometric analysis on T lymphocytes and natural killer cells revealed increased $T_H$/$T_S$ ratio and increased NK cells in red ginseng-treated group. These results suggest that the anticarcinogenic effect of red ginseng may be exerted by the increased cell-mediated immunity and natural killer cell activity.