• Journal of the Society of Cosmetic Scientists of Korea
• /
• v.48 no.1
• /
• pp.11-24
• /
• 2022

It has been reported that the ethanolic extract of the root of Piper methysticum (P. methysticum) inhibits melanogenesis in melanocyte stimulating hormone (MSH)-activated B16 melanoma cells. Flavokawain B (FKB) and Flavokawain C (FKC) isolated from this extract have been found to inhibit melanin production based on anti-melanogenesis activity. This study was designed to find out the inhibition and its process of FKB and FKC on melanin synthesis in melan-a melanocytes. FKB and FKC inhibited melanogenesis at 10 μM, 5 μM respectively in melan-a melanocytes. However, they did not inhibit extracellular tyrosinase activity from melan-a melanocytes. FKB reduced the protein level of tyrosinase (Tyr), tyrosinase-related protein 1 (TRP-1), tyrosinase-related protein 2 (TRP-2), microphthalmia-associated transcription factor (MITF) and the mRNA level of Tyr and TRP-1. FKC reduced the protein level of TRP-2 and MITF and the mRNA level of TRP-1 and Tyr. The reduced expression of Tyr and TRP-1 might be resulted from the decreased MITF which regulates major melanogenic proteins. However, since the mRNA expression of MITF did not change by FKB and FKC treatment, the effects of FKB and FKC on extracellular signal regulating kinase (ERK)/AKT phosphorylation, known to regulate the degradation of MITF, were confirmed. FKB and FKC significantly increased the phosphorylation of ERK1/2, not in AKT. These results suggest that FKB and FKC may be helpful as a potential depigmenting agent for various hyper-pigmentary disorders.

• Natural Product Sciences
• /
• v.17 no.1
• /
• pp.26-32
• /
• 2011

Novel tyrosinase inhibitors are important for pigmentation in the skin. Following extraction of tyrosinase inhibitors from edible vegetables or fruits, we found that the Prunus persica flesh extract (PPFE) exhibited potential inhibitory activity for melanogenesis. PPFE showed tyrosinase inhibitory activity in an enzymatic assay and PPFE also significantly inhibited the melanin formation in cultured mouse melan-a cells. Moreover, real-time RT-PCR analysis revealed that the inhibition of melanin production by PPFE was closely related to marked suppression of mRNA expression of tyrosinase and tyrosinase-related protein-1 and -2 (TRP-1 and TRP-2) in melan-a cells. Further investigation found that the modulation of tyrosinase expression by PPFE was associated with the transcriptional regulation of the microphthalmia-associated transcription factor (MITF). PPFE inhibited the promoter activity of MITF and suppressed MITF mRNA expression in melan-a cells. These results indicate that PPFE down-regulates melanogenesis-associated gene expression through MITF-mediated transcriptional regulation and these events might be related to the hypopigmentary effects of PPFE.

• Journal of Ginseng Research
• /
• v.41 no.3
• /
• pp.268-276
• /
• 2017

Background: UV-B-exposed keratinocytes secrete various paracrine factors. Among these factors, basic fibroblast growth factor (bFGF) stimulates the proliferation of melanocytes. Ginsenosides, the major active compounds of ginseng, are known to have broad pharmacological effects. In this study, we examined the antiproliferative effects of ginsenosides on bFGF-induced melanocyte proliferation. Methods: We investigated the inhibitory effects of Korean Red Ginseng and ginsenosides from Panax ginseng on bFGF-induced proliferation of melan-a melanocytes. Results: When melan-a melanocytes were treated with UV-B-irradiated SP-1 keratinocytes media, cell proliferation increased. This increased proliferation of melanocytes decreased with a neutralizing anti-bFGF antibody. To elucidate the effects of ginsenosides on melanocyte proliferation induced by bFGF, we tested 15 types of ginsenoside compounds. Among them, Rh3, Rh1, F1, and CK demonstrated antiproliferative effects on bFGF-induced melanocyte proliferation after 72 h of treatment. bFGF stimulated cell proliferation via extracellular signal-regulated kinase (ERK) activation in various cell types. Western blot analysis found bFGF-induced ERK phosphorylation in melan-a. Treatment with Rh3 inhibited bFGF-induced maximum ERK phosphorylation and F1-delayed maximum ERK phosphorylation, whereas Rh1 and CK had no detectable effects. In addition, cotreatment with Rh3 and F1 significantly suppressed bFGF-induced ERK phosphorylation. Western blot analysis found that bFGF increased microphthalmia-associated transcription factor (MITF) protein levels in melan-a. Treatment with Rh3 or F1 had no detectable effects, whereas cotreatment with Rh3 and F1 inhibited bFGF-induced MITF expression levels more strongly than a single treatment. Conclusion: In summary, we found that ginsenosides Rh3 and F1 have a synergistic antiproliferative effect on bFGF-induced melan-a melanocyte proliferation via the inhibition of ERK-mediated upregulation of MITF.

• Korean Journal of Medicinal Crop Science
• /
• v.17 no.4
• /
• pp.293-300
• /
• 2009

This study was carried out to investigate the effect of Cinnamomum camphora on melanogenesis. The MeOH extract of Cinnamomum camphora inhibited mushroom tyrosinase activity in dose-dependent manner. Moreover, it significantly suppressed the melanin production in melan-a cells at the concentration of $100{\mu}/m{\ell}$. The MeOH extract was partitioned with ethyl acetate, n-butanol and water. Among them, the BuOH soluble fraction exhibited significant inhibitory effect on mushroom tyrosinase. In addition, the BuOH soluble fraction reduced the melanin production in melan-a cells. But, the BuOH soluble fraction had less inhibition effects on melan-a cell originated tyrosinase. So, it was performed western blotting for melanogenic proteins (tyrosinase, tyrosinase-related protein (TRP-2)) using melan-a cells. The BuOH soluble fraction inhibited the protein expression of tyrosinase at the concentration of $100{\mu}/m{\ell}$. The results suggested that the BuOH soluble fraction of C. camphora might be a potent inhibitor of melanin biosynthesis in melan-a cells.

• Journal of the Korea Academia-Industrial cooperation Society
• /
• v.12 no.9
• /
• pp.4038-4045
• /
• 2011

To investigate whitening Effects of Angelica dahurica Radix Ethanol Extract (ADEE), we used melan-a cell line, brown guinea pig, and HMB-45. We treated with ADEE of 6.25, 12.5, 25, and 50 ${\mu}g/m{\ell}$ concentration in order to evaluate the effect of ADEE on cell viability and on morphological observation of melan-a cells. Also we were induced the artificial tanning spots by 1,500 mJ/$cm^2$ of ultraviolet B radiation on the backs of brown guinea pigs (approximately 450~500g) and then the test agent of $30{\mu}{\ell}$ was applied on the spots twice a day, five days a week, for five weeks respectively. The visible whitening effect was evaluated once a week. At the end of the experiment, the animals were sacrificed under anesthetization. The artificial tanning spots were obtained by biopsy punch and stained with HMB-45 to observe the gp100 proteins which were melanosomes. Our results show that cell viability was not reduce at ADEE concentrations between 6.25 and 50 ${\mu}g/m{\ell}$, melanin synthesis and melanocyte dendricity were decreased in ADEE treated melan-a cells increasing ADEE concentration. In the gross observation, ADEE treated groups had lower pigmentation than the vehicle control groups. And in the histological observation, ADEE treated groups had lower melanocytes than the vehicle control groups. Also in the quantitative analysis of the gp100 proteins using image analysis software, ADEE treated groups had a significantly lower value (p<0.001) than the vehicle control group and this resultsagreed with the results of observation under microscope. From these results, weconcluded that ADEE had positive whitening effect.

• Journal of Ginseng Research
• /
• v.41 no.4
• /
• pp.602-607
• /
• 2017

Background: Panax ginseng is a traditional herb used for medicinal purposes in eastern Asia. P. ginseng contains various ginsenosides with pharmacological effects. In this study, floralginsenoside A (FGA), ginsenoside Rd (GRD), and ginsenoside Re (GRE) were purified from P. ginseng berry. Methods: Chemical structures of FGA, GRD, and GRE were determined based on spectroscopic methods, including fast atom bombardment mass spectroscopy, ID-nuclear magnetic resonance, and infrared spectroscopy. Inhibitory activities of these compounds on melanogenesis were studied by measuring the expression of protein and melanin content in the melan-a cell line. This inhibitory activity was confirmed by observing pigmentation and tyrosinase activities of zebrafish. Results: GRD, GRE, and FGA were not cytotoxic at concentrations less than $20{\mu}M$, $80{\mu}M$, and $160{\mu}M$ in melan-a cells, respectively. GRD, GRE, and FGA inhibited melanin biosynthesis in melan-a cells by 15.2%, 22.9%, and 23.9% at $20{\mu}M$, $80{\mu}M$, and $160{\mu}M$, respectively. FGA was observed to display the most potent inhibitory effect. In addition, FGA decreased microphthalmia-associated transcription factor protein expression in a dose-dependent manner. Moreover, FGA induced extracellular signal-regulated kinase phosphorylation level in melan-a cells. In addition, melanin pigment content and tyrosinase activity in zebrafish treated with FGA at $160{\mu}M$ were reduced. Conclusion: FGA showed the most potent inhibition of melanogenesis in both in vitro and in vivo studies. This study suggests that FGA purified from P. ginseng may be an effective melanogenesis inhibitor.

• Toxicological Research
• /
• v.33 no.1
• /
• pp.55-62
• /
• 2017

We evaluated the antioxidant activity and anti-melanogenic effects of Oenothera laciniata methanol extract (OLME) in vitro by using melan-a cells. The total polyphenol and flavonoid content of OLME was 66.3 and 19.0 mg/g, respectively. The electron-donating ability, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical-scavenging activity, and superoxide dismutase (SOD)-like activity of OLME ($500{\mu}g/mL$) were 94.5%, 95.6%, and 63.6%, respectively. OLME and arbutin treatment at $50{\mu}g/mL$ significantly decreased melanin content by 35.5% and 14.2%, respectively, compared to control (p < 0.05). OLME and arbutin treatment at $50{\mu}g/mL$ significantly inhibited intra-cellular tyrosinase activity by 22.6% and 12.6%, respectively, compared to control (p < 0.05). OLME ($50{\mu}g/mL$) significantly decreased tyrosinase, tyrosinase-related protein-1 (TRP-1), TRP-2, and microphthalmia-associated transcription factor-M (MITF-M) mRNA expression by 57.1%, 67.3%, 99.0%, and 77.0%, respectively, compared to control (p < 0.05). Arbutin ($50{\mu}g/mL$) significantly decreased tyrosinase, TRP-1, and TRP-2 mRNA expression by 24.2%, 42.9%, and 48.5%, respectively, compared to control (p < 0.05). However, arbutin ($50{\mu}g/mL$) did not affect MITF-M mRNA expression. Taken together, OLME showed a good antioxidant activity and anti-melanogenic effect in melan-a cells that was superior to that of arbutin, a well-known skin-whitening agent. The potential mechanism underlying the anti-melanogenic effect of OLME was inhibition of tyrosinase activity and down-regulation of tyrosinase, TRP-1, TRP-2, and MITF-M mRNA expression.

• Toxicological Research
• /
• v.31 no.2
• /
• pp.165-172
• /
• 2015

Cornus officinalis is widely distributed in Korea, and its fruit has been used to make as herbal drug for traditional medicine in Korea, Japan, and China because of its tonic, analgesic, and diuretic properties. However, the effects of C. officinalis methanol extract (COME) on melanogenesis remain poorly understood. We evaluated the melanogenic capability of COME in melan-a cells, which are immortalized mouse melanocytes. COME increased melanin synthesis in a dose-dependent manner. Treatment with $12.5{\mu}g/mL$ of COME significantly increased melanin content by 36.1% (p < 0.001) to a level even higher than that (31.6%) of 3-isobutyl-1-methyl-xanthine, a well-known pigmentation agent. COME also upregulated tyrosinase activity and its messenger RNA and protein expression. In addition, COME upregulated the expression of tyrosinase-related proteins 1 and 2 and microphthalmia-associated transcription factor-M messenger RNA expression. These results imply that COME may be appropriate for development as a natural product to treat hair graying.

• Korean Journal of Veterinary Service
• /
• v.34 no.3
• /
• pp.273-278
• /
• 2011

Sertoli cell tumor (SCT) of the testicle arises from the supporting cells within the seminiferous tubules. SCT is common in dogs, especially in cryptorchid testicles, but also has been reported in the stallion, ram, cat, and bull. Sertoli cell tumor sample was collected from 7-years male german shepherd. In this study, SCT arose from one testicle. Sample size is approximately 1.7 cm in diameter and it has a round form. In the microscopic, cells within the tumor variably resemble Sertoli cells (SCs) that normally populate the seminiferous tubules and interstitial area. There is abundant stroma of dense, mature fibrous connective tissue in SCT. In the immunohistochemical staining, cytokeratin AE1/AE3 was not expressed in the control and SCT. S-100 protein was expressed by SCs, germ cells and fibrous connective tissue of SCT. Melan A was expressed by leydig cells (LCs) of SCT. A study by using S-100 and melan A in canine SCT was almost never carried out. S-100 and melans A is considered to suggest for diagnosis and pathogenesis of canine SCTs. Inhibin-alpha and Vimentin were well known as the marekers of SCTs. Also, they were expressed by Sertoli cells and LSs of SCT in this study.

• Journal of Microbiology and Biotechnology
• /
• v.32 no.8
• /
• pp.982-988
• /
• 2022

Licorice (Glycyrrhiza) has been used as preventive and therapeutic material for hyperpigmentation disorders. Previously, we isolated noble compounds including dehydroglyasperin C (DGC), dehydroglyasperin D (DGD) and isoangustone A (IAA) from licorice hexane/ethanol extracts. However, their anti-melanogenic effects and underlying molecular mechanisms are unknown. The present study compared effects of DGC, DGD and IAA on pigmentation in melan-a melanocytes and human epidermal melanocytes (HEMn). DGD exerted the most excellent anti-melanogenic effect, followed by DGC and IAA at non-cytotoxic concentrations. In addition, DGD significantly inhibited tyrosinase activity in vitro cell-free system and cell system. Western blot result showed that DGD decreased expression of microphthalmia-associated transcription factor (MITF), tyrosinase and tyrosinase-related protein-1 (TRP-1) in melan-a cells and HEMn cells. DGD induced phosphorylation of MITF, ERK and Akt signal pathway promoting MITF degradation system. However, DGD did not influence p38 and cAMP-dependent protein kinase (PKA)/CREB signal pathway in melan-a cells. These result indicated that DGD inhibited melanogenesis not only direct regulation of tyrosinase but also modulating intracellular signaling related with MITF level. Collectively, these results suggested a protective role for DGD against melanogenesis.