Inhibitory Effects of Pinostilbene Hydrate on Melanogenesis in B16F10 Melanoma Cells via ERK and p38 Signaling Pathways

Melanin protects our skin from harmful ultraviolet (UV) radiation. However, when produced in excess, it can cause hyperpigmentation disorders, such as melanoma, freckles, lentigo, and blotches. In this study, we investigated the effects of pinostilbene hydrate (PH) on melanogenesis. We also examined the underlying mechanisms of PH on melanin production in B16F10 cells. Our findings indicated that PH significantly inhibits melanin content and cellular tyrosinase activity in cells without causing cytotoxicity. In addition, Western blot analysis showed that PH downregulated the protein levels of microphthalmia-associated transcription factor (MITF), tyrosinase, and other melanogenic enzymes, such as tyrosinase-related protein-1 (TRP-1) and tyrosinase-related protein-2 (TRP-2). Although PH activated the phosphorylation of extracellular signal-regulated kinase (ERK), it inhibited p38 mitogen-activated protein kinases (p38). Furthermore, the inhibition of tyrosinase activity by PH was attenuated by treatment with PD98059 (a specific ERK inhibitor). Additionally, p-AKT was upregulated by PH treatment. Finally, the inhibitory effects of PH on melanin content and tyrosinase activity were confirmed in normal human melanocytes. These results suggest PH downregulates melanogenesis via the inhibition of MITF expression, followed by the MAPKase signaling pathways. Thus, PH may be used to treat or prevent hyperpigmentation disorders and in functional cosmetic agents for skin whitening.

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Inhibitory Effects of Cycloheterophyllin on Melanin Synthesis

Molecules ◽

10.3390/molecules26092526 ◽

2021 ◽

Vol 26 (9) ◽

pp. 2526

Author(s):

Joong-Hyun Shim

Keyword(s):

Functional Materials ◽

Tyrosinase Activity ◽

Melanin Synthesis ◽

Activity Assay ◽

Inhibitory Effects ◽

Melanin Production ◽

Messenger Ribonucleic Acid ◽

B16f10 Cells ◽

Cell Line Cell ◽

Tyrosinase Related Protein

This study was performed to clarify the inhibitory effects of cycloheterophyllin on melanin synthesis. In order to elucidate the inhibitory effects of cycloheterophyllin on the B16F10 cell line, cell viability, messenger ribonucleic acid (mRNA) expressions, tyrosinase activity assay, and melanin production assay were measured. The effects of cycloheterophyllin on tyrosinase-related protein 1 (TYRP1)/TYRP2/tyrosinase (TYR)/microphthalmia-associated transcription factor (MITF) mRNA expressions and melanin content were determined. Quantitative real-time RT-PCR showed that cycloheterophyllin decreased the mRNA expression level of TYRP1/TYRP2/TYR/MITF genes and melanin production contents than α-MSH-treated B16F10 cells. The tyrosinase activity assay revealed that cycloheterophyllin decreased the melanin production in the B16F10 cells. These data show that cycloheterophyllin increases the whitening effects in the B16F10 cells; thus, cycloheterophyllin is a potent ingredient for skin whitening. Thus, further research on the mechanism of action of cycloheterophyllin for the development of functional materials should be investigated.

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Anti-Melanogenic Effects of Hydroxyectoine via MITF Inhibition by JNK, p38, and AKT Pathways in B16F10 Melanoma Cells

Natural Product Communications ◽

10.1177/1934578x19858523 ◽

2019 ◽

Vol 14 (6) ◽

pp. 1934578X1985852 ◽

Cited By ~ 1

Author(s):

You C. Chung ◽

Min-Jin Kim ◽

Eun Y. Kang ◽

Yun B. Kim ◽

Bong S. Kim ◽

...

Keyword(s):

Skin Color ◽

Inhibitory Effect ◽

Tyrosinase Activity ◽

Dependent Manner ◽

Related Protein ◽

B16f10 Melanoma ◽

Melanin Production ◽

B16f10 Cells ◽

Dose Dependent ◽

Tyrosinase Related Protein

Melanin plays a role in determining human skin color of a person, and a large amount of melanin makes the skin color look darkened. The proper amount of melanin formation protects our skin from UV radiation, but excessive melanin production causes hyperpigmentation and leads to freckles, melasma, and lentigo. In this study, we investigated the inhibitory effect of hydroxyectoine on melanogenesis and its mechanism in B16F10 cells. Melanin content and cellular tyrosinase activity were determined. The expression of microphthalmia-associated transcription factor (MITF), and the activities of tyrosinase and other melanogenesis-related enzymes, such as tyrosinase-related protein 1 (TRP-1) and tyrosinase-related protein 2, were also examined. Hydroxyectoine treatment significantly inhibited melanin production and intracellular tyrosinase activity in a dose-dependent manner. Western blot analysis showed that hydroxyectoine also reduced the expressions of tyrosinase and TRP-1. In addition, hydroxyectoine significantly reduced the expression of MITF, a major regulator of melanin production, and inhibited the phosphorylation of p38, c-Jun N-terminal kinase, and activated the protein kinase B. The results demonstrated that hydroxyectoine inhibits the expression of MITF through the inhibition or activation of melanin-related signaling pathways and downregulates melanogenesis by inhibiting melanogenic enzyme expression and tyrosinase activity. Hydroxyectoine has potential value in functional cosmetics applications, such as whitening.

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Induction of Melanogenesis by Fosfomycin in B16F10 Cells Through the Upregulation of P-JNK and P-p38 Signaling Pathways

Antibiotics ◽

10.3390/antibiotics9040172 ◽

2020 ◽

Vol 9 (4) ◽

pp. 172

Author(s):

Sana Ullah ◽

You Chul Chung ◽

Chang-Gu Hyun

Keyword(s):

Western Blotting ◽

Disodium Salt ◽

Water Soluble ◽

Dependent Manner ◽

Related Protein ◽

Melanin Content ◽

B16f10 Cells ◽

Tract Infections ◽

Dose Dependent ◽

Tyrosinase Related Protein

Fosfomycin disodium salt (FDS), which is a water-soluble extract, is a bactericidal drug used to inhibit the synthesis of cells. Moreover, it has been found to be effective in the treatment of urinary tract infections. The present study was conducted to investigate the melanogenesis-stimulating effect of FDS in B16F10 cells. Several experiments were performed on B16F10 cells: the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, the melanin content assay, the cellular tyrosinase activity assay, and Western blotting. FDS upregulated the activity of tyrosinase in a dose-dependent manner at a wide concentration range of 0–1 mg/mL, which showed no cytotoxicity. It also increased the melanin content and the activity of the microphthalmia-associated transcription factor (MITF), tyrosinase related protein 1 (TRP-1), and tyrosinase related protein 2 (TRP-2) enzymes in a dose-dependent manner. Western blotting results showed that FDS clearly upregulated the phosphorylation of c-Jun N-terminal kinases (JNK) and p38 pathways. These data are clear evidence of the melanogenesis-inducing effect of FDS in B16F10 murine melanoma cells.

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Phoenix dactylifera L. Seed Extract Exhibits Antioxidant Effects and Attenuates Melanogenesis in B16F10 Murine Melanoma Cells by Downregulating PKA Signaling

Antioxidants ◽

10.3390/antiox9121270 ◽

2020 ◽

Vol 9 (12) ◽

pp. 1270

Author(s):

Huey-Chun Huang ◽

Shr-Shiuan Wang ◽

Tsang-Chi Tsai ◽

Wang-Ping Ko ◽

Tsong-Min Chang

Keyword(s):

Phoenix Dactylifera ◽

Seed Extract ◽

Skin Care ◽

Tyrosinase Activity ◽

Total Phenolic ◽

Related Protein ◽

Ros Scavenging ◽

B16f10 Cells ◽

Antioxidant Characteristics ◽

Tyrosinase Related Protein

Background: The mode of action of Phoenix dactylifera seed extract in skin care has never been explored. Methods: P. dactylifera L. seeds were extracted by ultrasonic extraction. The antioxidant characteristics of the extract were determined by 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-di-(3-ethylbenzthiazoline sulfonic acid) (ABTS+) assays and scavenging methods. The total phenolic content, reducing capacity, iron (II) ion-chelation, and intracellular reactive oxygen species (ROS)-scavenging capacities were also investigated. The effects of P. dactylifera L. seed extract on melanogenesis were evaluated spectrophotometrically by a mushroom tyrosinase activity assay, determination of intracellular tyrosinase activity, and melanin content. The expression levels of melanogenesis-related proteins were analyzed by Western blotting. Results: The results revealed that the P. dactylifera L. seed extract exerted apparent antioxidant capacity and significantly decreased intracellular ROS content at concentrations of 0.245 and 0.49 (mg/mL). Furthermore, the extract decreased the expression of melanocortin 1 receptor (MC1R), microphthalmia-associated transcription factor (MITF), tyrosinase, tyrosinase-related protein-1 (TRP1), and tyrosinase-related protein-2 (TRP2), and inhibited melanogenesis in B16F10 cells. Conclusions: Our results revealed that P. dactylifera L. seed extract attenuated melanogenesis in B16F10 cells by downregulating protein kinase A (PKA) signaling pathways. Hence, the extract could be used as a type of skin-whitening agent in skin care products.

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Exploring the potential effect and mechanisms of protocatechuic acid on human hair follicle melanocytes

Acta Pharmaceutica ◽

10.2478/acph-2020-0023 ◽

2020 ◽

Vol 70 (4) ◽

pp. 539-549

Author(s):

Bo Li ◽

Jun Tan ◽

Bosheng Zou ◽

Xiaojia Liu ◽

Yiling Yu

Keyword(s):

Hair Follicle ◽

Human Hair ◽

Protocatechuic Acid ◽

Treatment Time ◽

Tyrosinase Activity ◽

Dependent Manner ◽

Related Protein ◽

Human Hair Follicle ◽

Dose Dependent ◽

Tyrosinase Related Protein

AbstractThis study aims to evaluate the effect of protocatechuic acid (PCA) on human hair follicle melanocytes (HFM). Normal primary HFM were isolated and cultured till logarithmic period of second passage, then treated with different concentrations of PCA (0.1–200 μmol L−1) to study the cell proliferation, melanin contents, tyrosinase activity and protein and mRNA expression of melanogenic genes (tyrosinase-related protein 1 (TRP-1), tyrosinase-related protein 2 (TRP-2), and microphthalmia-associated transcription factor (MITF)) in the cultured HFM. In addition, we have also measured the contents of superoxide dismutase (SOD) and glutathione (GSH) in PCA treated HFM. Vitamin C was used as a positive control. The result showed that PCA can decrease the synthesis of melanin and the tyrosinase activity with IC50 = 8.9 μmol L−1 and IC50 = 6.4 μmol L−1, respectively, at the treatment time of 24 hours, without inducing any cytotoxicity in HFM cells. In addition, the mRNA transcription and protein expression levels of TRP-1, TRP-2 and MITF significantly decreased with a dose-dependent manner after 24-hour PCA treated in HFM cells. Furthermore, PCA has significantly increased the SOD and GSH activity in a dose-dependent manner for 24-hour PCA treatment. This study suggested that PCA has an inhibitory effect on the production of melanin through down-regulation of the expression of melanogenesis-related protein and the effect of anti-oxidation, which could be useful for the therapy of melanin overproduction or skin whitening.

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Praeruptorin B Mitigates the Metastatic Ability of Human Renal Carcinoma Cells through Targeting CTSC and CTSV Expression

International Journal of Molecular Sciences ◽

10.3390/ijms21082919 ◽

2020 ◽

Vol 21 (8) ◽

pp. 2919

Author(s):

Chia-Liang Lin ◽

Tung-Wei Hung ◽

Tsung-Ho Ying ◽

Chi-Jui Lin ◽

Yi-Hsien Hsieh ◽

...

Keyword(s):

Growth Factor Receptor ◽

Cellular Migration ◽

Mitogen Activated Protein Kinase ◽

Migration And Invasion ◽

Antitumor Effects ◽

Protein Levels ◽

Extracellular Signal ◽

Adult Kidney ◽

Mitogen Activated Protein ◽

Underlying Mechanisms

Renal cell carcinoma (RCC) is the most common adult kidney cancer, and accounts for 85% of all cases of kidney cancers worldwide. Praeruptorin B (Pra-B) is a bioactive constituent of Peucedanum praeruptorum Dunn and exhibits several pharmacological activities, including potent antitumor effects. However, the anti-RCC effects of Pra-B and their underlying mechanisms are unclear; therefore, we explored the effects of Pra-B on RCC cells in this study. We found that Pra-B nonsignificantly influenced the cell viability of human RCC cell lines 786-O and ACHN at a dose of less than 30 μM for 24 h treatment. Further study revealed that Pra-B potently inhibited the migration and invasion of 786-O and ACHN cells, as well as downregulated the mRNA and protein expression of cathepsin C (CTSC) and cathepsin V (CTSV) of 786-O and ACHN cells. Mechanistically, Pra-B also reduced the protein levels of phospho (p)-epidermal growth factor receptor (EGFR), p-mitogen-activated protein kinase kinase (MEK), and p-extracellular signal-regulated kinases (ERK) in RCC cells. In addition, Pra-B treatment inhibited the effect of EGF on the upregulation of EGFR–MEK–ERK, CTSC and CTSV expression, cellular migration, and invasion of 786-O cells. Our findings are the first to demonstrate that Pra-B can reduce the migration and invasion ability of human RCC cells through suppressing the EGFR-MEK-ERK signaling pathway and subsequently downregulating CTSC and CTSV. This evidence suggests that Pra-B can be developed as an effective antimetastatic agent for the treatment of RCC.

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Melanocytes and Pigmentation Are Affected in Dopachrome Tautomerase Knockout Mice

Molecular and Cellular Biology ◽

10.1128/mcb.24.8.3396-3403.2004 ◽

2004 ◽

Vol 24 (8) ◽

pp. 3396-3403 ◽

Cited By ~ 75

Author(s):

Laurence Guyonneau ◽

Fabien Murisier ◽

Anita Rossier ◽

Alexandre Moulin ◽

Friedrich Beermann

Keyword(s):

Knockout Mice ◽

Null Allele ◽

Coat Color ◽

Pigment Epithelium ◽

Retinal Pigment ◽

Related Protein ◽

Melanin Content ◽

Dopachrome Tautomerase ◽

Exon 1 ◽

Tyrosinase Related Protein

ABSTRACT The tyrosinase family comprises three members, tyrosinase (Tyr), tyrosinase-related protein 1 (Tyrp1), and dopachrome tautomerase (Dct). Null mutations and deletions at the Tyr and Tyrp1 loci are known and phenotypically affect coat color due to the absence of enzyme or intracellular mislocalization. At the Dct locus, three mutations are known that lead to pigmentation phenotype. However, these mutations are not null mutations, and we therefore set out to generate a null allele at the Dct gene locus by removing exon 1 of the mouse Dct gene. Mice deficient in Dct [Dcttm1(Cre)Bee ] lack Dct mRNA and dopachrome tautomerase protein. They are viable and do not show any abnormalities in Dct-expressing sites such as skin, retinal pigment epithelium, or brain. However, the mice show a diluted coat color phenotype, which is due to reduced melanin content in hair. Primary melanocytes from Dct knockout mice are viable in culture and show a normal distribution of tyrosinase and tyrosinase-related protein 1. In comparison to the knockout, the slaty mutation (Dctslt /Dctslt ) has less melanin and affects growth of primary melanocytes severely. In summary, we have generated a knockout of the Dct gene in mice with effects restricted to pigment production and coat color.

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Mitogen-activated protein kinase activation and regulation in the pressure-loaded fetal ovine heart

AJP Heart and Circulatory Physiology ◽

10.1152/ajpheart.00984.2005 ◽

2006 ◽

Vol 290 (4) ◽

pp. H1587-H1595 ◽

Cited By ~ 11

Author(s):

Aaron K. Olson ◽

Kristin N. Protheroe ◽

Jeffrey L. Segar ◽

Thomas D. Scholz

Keyword(s):

Map Kinase ◽

Signaling Pathways ◽

Fetal Heart ◽

Left Ventricular ◽

Heart Weight ◽

Kinase Signaling ◽

Protein Levels ◽

Pressure Load ◽

Map Kinase Signaling ◽

Mitogen Activated Protein

The mitogen-activated protein (MAP) kinase signaling pathways help to mediate the hypertrophic response of the pressure-loaded adult heart, although their importance in fetal myocardium is less known. The goal of this study was to determine the role the MAP kinase signaling pathways play in regulating the response of the fetal heart to a pressure load. Aortic (Ao) and pulmonary artery (PA) bands were placed in 132-day fetal sheep for 7 days. Protein levels of the total and active (phosphorylated) terminal MAP kinases extracellular signal-regulated kinase (ERK/P-ERK), c-Jun NH2-terminal kinase (JNK/P-JNK), and p38/P-p38 and the MAP kinase phosphatases MKP-1, MKP-2, and MKP-3 were made in the right and left ventricular (RV and LV) free walls. In both Ao- and PA-banded animals, total heart weight normalized to body weight was significantly increased, largely due to an increase in RV free wall mass in the Ao-banded animals and an increase in septal mass in the PA-banded fetuses. Total protein levels of the three terminal kinases and of P-ERK and P-JNK remained stable in both groups of banded animals. However, P-p38 was significantly increased in RV and LV of Ao- and PA-banded fetuses. Whereas MKP-1 and MKP-2 protein levels were unchanged following Ao- and PA-banding, MKP-3 protein levels were significantly increased in the RV of the PA-banded animals. These findings indicate that the MAP kinase signaling pathways are active in the fetal heart and help to modulate the response of prenatal myocardium to a pressure load.

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Role of EGF receptor tyrosine kinase activity in antiapoptotic effect of EGF on mouse hepatocytes

AJP Gastrointestinal and Liver Physiology ◽

10.1152/ajpgi.2001.280.6.g1360 ◽

2001 ◽

Vol 280 (6) ◽

pp. G1360-G1369 ◽

Cited By ~ 34

Author(s):

Lina Musallam ◽

Chantal Éthier ◽

Pierre S. Haddad ◽

Marc Bilodeau

Keyword(s):

Tyrosine Kinase ◽

Egf Receptor ◽

Phosphatidylinositol 3 Kinase ◽

Antiapoptotic Effect ◽

Protein Levels ◽

Primary Mouse ◽

Mitogen Activated Protein ◽

Epidermal Growth ◽

Mouse Hepatocytes ◽

Underlying Mechanisms

The apoptotic Fas pathway is potentially involved in the pathogenesis of liver diseases. Growth factors, such as epidermal growth factor (EGF), can protect cells against apoptosis induced by a variety of stimuli, including Fas receptor stimulation. However, the underlying mechanisms of this protection have yet to be determined. We investigated the involvement of EGF receptor (EGFR) tyrosine kinase (TK) activity in the antiapoptotic effect of EGF on primary mouse hepatocyte cultures. Cells undergoing apoptosis after treatment with anti-Fas antibody were protected by EGF treatment. This protection was significantly but partially decreased when cells were treated with two specific inhibitors of the TK activity of EGFR. Evaluation of the efficacy of these compounds indicated that they were able to abolish EGFR autophosphorylation and postreceptor events such as activation of mitogen-activated protein kinases and the phosphatidylinositol 3′-kinase pathways as well as increases in Bcl-xL mRNA and protein levels. This leads us to postulate that EGF exerts its antiapoptotic action partially through the TK activity of EGFR. In addition, our results suggest that Bcl-xL protein upregulation caused by EGF is linked to the TK activity of its receptor.

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Liquiritin and Liquiritigenin Induce Melanogenesis via Enhancement of p38 and PKA Signaling Pathways

Medicines ◽

10.3390/medicines6020068 ◽

2019 ◽

Vol 6 (2) ◽

pp. 68 ◽

Cited By ~ 1

Author(s):

Takuhiro Uto ◽

Tomoe Ohta ◽

Akihisa Yamashita ◽

Shunsuke Fujii ◽

Yukihiro Shoyama

Keyword(s):

Signaling Pathways ◽

Mek Inhibitor ◽

Tyrosinase Activity ◽

Licorice Root ◽

Melanin Synthesis ◽

Creb Phosphorylation ◽

Pi3k Inhibitor Ly294002 ◽

P38 Inhibitor ◽

Protein Levels ◽

Mitf Expression

Background: Liquiritin (LQ) and its aglycone, liquiritigenin (LQG), are major flavonoids in licorice root (Glycyrrhiza spp.). Our preliminary screening identified LQ and LQG, which promote melanin synthesis in the melanoma cells. In this study, we investigated the molecular mechanism of melanin synthesis activated by LQ and LQG. Methods: Murine (B16-F1) and human (HMVII) melanoma cell lines were treated with LQ or LQG. After incubation, melanin contents, intracellular tyrosinase activity, and cell viability were evaluated. Protein levels were determined using Western blotting. Results: LQ and LQG activated melanin synthesis and intracellular tyrosinase activity. The induction of melanin and intracellular tyrosinase activity by LQG was higher than that by LQ. LQ and LQG induced the expression of tyrosinase, tyrosinase-related protein (TRP)-1, and TRP-2. LQ and LQG also enhanced microphthalmia-associated transcription factor (MITF) expression, and cyclic AMP-responsive element-binding protein (CREB) phosphorylation. The phosphorylation of p38 and extracellular signal-regulated kinase (ERK), but not Akt, was significantly increased by LQ or LQG. Furthermore, LQ- or LQG-mediated melanin synthesis was partially blocked by p38 inhibitor (SB203580) and protein kinase A (PKA) inhibitor (H-89); however, ERK kinase (MEK) inhibitor (U0126) and phosphatidylinositol-3-kinase (PI3K) inhibitor (LY294002) had no effect. Conclusions: The results suggest that LQ and LQG enhance melanin synthesis by upregulating the expression of melanogenic enzymes, which were activated by p38 and PKA signaling pathways, leading to MITF expression and CREB phosphorylation.

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