Pterostilbene Inhibits the Melanogenesis Activity in UVB-Irradiated B164A5 Cells

Pterostilbene is a potent antioxidant and anti-inflammatory agent. However, its chemopreventive effects via anti-tyrosinase activity and inhibitory effects on melanin content have not been reported previously. Hence, this study aimed to investigate the anti-melanogenic activity of pterostilbene on UVB-irradiated B164A5 mouse melanoma cells. The effects of pterostilbene and resveratrol on cell viability were determined by MTT assay, whereas melanin content and tyrosinase assay were employed to assess melanogenesis activity. Western blot analysis was performed to determine the tyrosinase expression. Based on the MTT assay, the IC50 value of pterostilbene on UVB-irradiated B164A5 cells was 34.0 ± 3.43 μM, in comparison to resveratrol (>100 μM). Next, 5 and 10 μM pterostilbene showed a significant dose-dependent inhibition ( P < .01) of tyrosinase activity in UVB-irradiated B164A5 cells at 37.14 ± 2.71% and 58.36 ± 6.8%, respectively. The findings from the tyrosinase assay also confirmed the downregulation of tyrosinase expression in UVB-irradiated B164A5 cells as measured by Western blot analysis. Finally, 10 μM pterostilbene showed a significantly decreased melanin content ( P < .01) in UVB-irradiated B164A5 cells, at 27.34 ± .98 μg/mL. In conclusion, pterostilbene showed anti-melanogenic activity that was 10 times more potent than resveratrol in the UVB-irradiated B164A5 cell.

Anti-Melanogenesis Activity of 6-O-Isobutyrylbritannilactone from Inula britannica on B16F10 Melanocytes and In Vivo Zebrafish Models

A potential natural melanogenesis inhibitor was discovered in the form of a sesquiterpene isolated from the flowers of Inula britannica, specifically 6-O-isobutyrylbritannilactone (IBL). We evaluated the antimelanogenesis effects of IBL on B16F10 melanocytes and zebrafish embryos. As a result, we found that 3-isobutyl-1-methylxanthine (IBMX)-induced melanin production was reduced in a dose-dependent manner in B16F10 cells by IBL. We also analyzed B16F10 cells that were and were not treated with IBMX, investigating the melanin concentration, tyrosinase activity, mRNA levels. We also studied the protein expressions of microphthalmia-associated transcription factor (MITF), tyrosinase, and tyrosinase-related proteins (TRP1, and TRP2). Furthermore, we found that melanin synthesis and tyrosinase expression were also inhibited by IBL through the modulation of the following signaling pathways: ERK, phosphoinositide 3-kinase (PI3K)/AKT, and CREB. In addition, we studied antimelanogenic activity using zebrafish embryos and found that the embryos had significantly reduced pigmentation in the IBL-treated specimens compared to the untreated controls.

Luteolin 7-Sulfate Attenuates Melanin Synthesis through Inhibition of CREB- and MITF-Mediated Tyrosinase Expression

Antioxidants with antimelanogenic activity are potentially useful for the attenuation of skin hyperpigmentation disorders. In a previous study, luteolin 7-sulfate isolated from Phyllospadix iwatensis Makino, a marine plant, was shown to inhibit cellular melanin synthesis. The aim of the present study was to examine its action mechanism, focusing on the regulation of tyrosinase (TYR) expression in cells. Cell-based assay was undertaken using murine melanoma B16-F10 cells and primary human epidermal melanocytes (HEMs). Luteolin 7-sulfate showed lower toxicity compared to luteolin in B16-F10 cells. At the non-toxic concentration ranges, luteolin 7-sulfate attenuated melanin synthesis, stimulated by α-melanocyte-stimulating hormone or forskolin. Luteolin 7-sulfate attenuated forskolin-induced microphthalmia-associated transcription factor (MITF) and TYR expressions at the mRNA and protein levels in B16-F10 cells. It also attenuated the phosphorylation of cAMP-responsive element binding protein (CREB) stimulated by forskolin. Luteolin 7-sulfate also attenuated melanin synthesis in primary HEMs. This study demonstrates that luteolin 7-sulfate attenuates TYR gene expression through the intervention of a CREB- and MITF-mediated signaling pathway, leading to the decreased melanin synthesis.

Conjugation with Dihydrolipoic Acid Imparts Caffeic Acid Ester Potent Inhibitory Effect on Dopa Oxidase Activity of Human Tyrosinase

Caffeic acid derivatives represent promising lead compounds in the search for tyrosinase inhibitors to be used in the treatment of skin local hyperpigmentation associated to an overproduction or accumulation of melanin. We recently reported the marked inhibitory activity of a conjugate of caffeic acid with dihydrolipoic acid, 2-S-lipoylcaffeic acid (LCA), on the tyrosine hydroxylase (TH) and dopa oxidase (DO) activities of mushroom tyrosinase. In the present study, we evaluated a more lipophilic derivative, 2-S-lipoyl caffeic acid methyl ester (LCAME), as an inhibitor of tyrosinase from human melanoma cells. Preliminary analysis of the effects of LCAME on mushroom tyrosinase indicated more potent inhibitory effects on either enzyme activities (IC50 = 0.05 ± 0.01 μM for DO and 0.83 ± 0.09 μM for TH) compared with LCA and the reference compound kojic acid. The inhibition of DO of human tyrosinase was effective (Ki = 34.7 ± 1.1 μM) as well, while the action on TH was weaker. Lineweaver–Burk analyses indicated a competitive inhibitor mechanism. LCAME was not substrate of tyrosinase and proved nontoxic at concentrations up to 50 μM. No alteration of basal tyrosinase expression was observed after 24 h treatment of human melanoma cells with the inhibitor, but preliminary evidence suggested LCAME might impair the induction of tyrosinase expression in cells stimulated with α-melanocyte-stimulating hormone. All these data point to this compound as a valuable candidate for further trials toward its use as a skin depigmenting agent. They also highlight the differential effects of tyrosinase inhibitors on the human and mushroom enzymes.

Fermented Onions Extract Inhibits Tyrosinase and Collagenase-1 Activities as a Potential New Anti–Photoaging Agent

The bulbs of onion ( Allium cepa L.) have long been used as food and dietary supplements in the treatment of various diseases. This study aimed to investigate anti-photoaging activities of fermented onions extract rich in flavonoids such as quercetin and isoquercetin. The fermented onions extract (FOE) effectively suppressed the melanin production by inhibition of tyrosinase expression in B16F10 melanoma cells at a concentration of 100 μg/mL. Besides, the FOE exhibited down-regulation of collagenase-1 expression and up-regulation of type I collagen level in UVB-irradiated HaCaT keratinocyte cells. Hyaluronic acid production was increased to 41.11% (10 μg/mL), 107.78% (100 μg/mL), and 146.67% (200 μg/mL) compared with the UVB-irradiated control by treatment of FOE. Therapeutic attempts with FOE might be useful in preventing or treating melanin pigmentary diseases and UVB-induced wrinkle formation.