Investigation of the Regulation of Pigmentation in α‐Melanocyte‐Stimulating Hormone Responsive and Unresponsive Cultured B16 Melanoma Cells

Hill, Sarah E.; Buffey, J.; Thody, A. J.; Oliver, Ian; Bleehen, S.S.; Neil, S. Mac

doi:10.1111/j.1600-0749.1989.tb00181.x

Cited by 44 publications

(34 citation statements)

References 19 publications

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“…Thus, we evaluated melanogenesis activity on the basis of the sum of the amount of intracellular and extracellular melanin. According to the method of Hill et al, 13) the amounts of intracellular and extracellular melanin were determined separately. The sum of the amount of melanin was calculated by the addition of both average melanin amounts, and was represented without statistical standard errors.…”

Section: Resultsmentioning

confidence: 99%

“…The results concerning the melanogenesis activities of sixteen coumarins (1)(2)(3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16) are shown in Fig. 3.…”

Section: Discussionmentioning

confidence: 99%

“…The amount of melanin in cultured B16 murine melanoma cells was measured according to the method of Hill et al 13) as described in one of our previous papers.…”

Section: Measurement Of Produced Melanin In Cultured B16 Melanoma Cellsmentioning

confidence: 99%

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Melanogenesis Stimulation in Murine B16 Melanoma Cells by Umberiferae Plant Extracts and Their Coumarin Constituents

Matsuda

Hirata

Kawaguchi

et al. 2005

Biological & Pharmaceutical Bulletin

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Gray hair is caused by a genetic predisposition, aging, decrement of melanocytes by environmental stress, and decrement of the biosynthesis of melanin pigment or melanogenesis.1,2) Hair dye agents are used for the treatment of gray hair. However, there remain some problems with these agents, such as side effects due to the dyes. Thus, there is a need for anti-gray hair agents that exhibit satisfactory melanogenesis activity and gray hair prevention. For the development of gray hair prevention agents, we have carried out a screening program to find a potential stimulant of melanogenesis from natural resources by using cultured murine B16 melanoma cells with theophylline as a reference drug. In a preceding paper, 3) we reported that a methanolic extract from the leaves of Piper nigrum L. showed a significant stimulatory effect on melanogenesis in cultured murine B16 melanoma cells and that two lignans, (Ϫ)-cubebin and (Ϫ)-3,4-dimethoxy-3,4-desmethylenedioxycubebin, were isolated as its active constituents. As the next step in our herbal screening, we report here the melanogenesis stimulation effect of seven ethanolic extracts obtained from Umbelliferae -dokkatsu), and the fruits of Cinidium monnieri (L.) CUSSON (Jashoshi) and C. formosanum YABE (Taiwan-jashoshi). The melanogenesis stimulation effects of sixteen coumarins (1-16, Fig. 1) isolated from the Umberiferae crude drugs were also evaluated. MATERIALS AND METHODS MaterialsThe roots of Angelica biserrata (Todokkatsu), Notopterygium incisum (Kyokatsu), the fruits of Cinidium monnieri (Jashoshi) and Cinidium formosanum (Taiwan-jashoshi) were obtained from Osaka market. The roots of Angelica dahurica (Byakushi) and Heracleum candicans (Hakuryo-dokkatsu) were obtained from Chinese market (Yunnan Province). The roots of Heracleum lanatum (Gyubi-dokkatsu) were grown at the Osaka University of Pharmaceutical Sciences.Preparation of Ethanolic Extracts Seven crude drugs (each 10 g) were extracted with EtOH (100 ml) in a water bath at 70°C for 2 h and then filtrated. Each solution was concentrated under reduced pressure to give the corresponding extract: To-dokkatsu (1.00 g), Kyokatsu (1.45 g), Byakushi (0.70 g), Hakuryo-dokkatsu (1.00 g), Gyubidokkatsu (1.17 g), Jashoshi (0.40 g), and Taiwan-jashoshi (0.40 g).Drugs Fourteen coumarins (1-7, 9, 11-16, Fig. 1) were isolated from the Umbelliferae crude drugs.4-10) The isolation yields of each compound are depicted in Table 1. Synthetic melanin and 0.4 M HEPES buffer (pH 6.8) were purchased from Sigma-Aldrich Japan (Tokyo, Japan). Dulbecco's modified Eagle medium (D-MEM), Antibiotic-Antimycotic (a mixture of 10000 U/ml penicillin, 10000 mg/ml streptomycin sulfate, and 25 mg/ml amphotericine B), Dulbecco's phosphate buffered saline (Ca Isolation of Heraclenin (8) and Heraclenol (10) from Hakuryo-dokkatsu (H. candicans WALL.) Powdered Hakuryo-dokkatsu (500 g) was successively extracted with hexane (5 l) and EtOAc (5 l) under reflux for 2 h. Each extract was evaporated under reduced pressure to give a hexane ex-

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Section: Resultsmentioning

confidence: 99%

“…The results concerning the melanogenesis activities of sixteen coumarins (1)(2)(3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16) are shown in Fig. 3.…”

Section: Discussionmentioning

confidence: 99%

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Melanogenesis Stimulation in Murine B16 Melanoma Cells by Umberiferae Plant Extracts and Their Coumarin Constituents

Matsuda

Hirata

Kawaguchi

et al. 2005

Biological & Pharmaceutical Bulletin

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“…The amount of melanin in cultured murine B16 melanoma cells (intracellular melanin) was measured according to the method of Hill et al, 18) as described in our previous paper. 19) Briefly, test samples were dissolved in DMSO/Ca Assay of Cell Proliferation The cell proliferation of murine B16 melanoma cells was assessed using the MTT method described in our previous paper.…”

Section: Measurement Of Melanin In Cultured B16 Melanoma Cellsmentioning

confidence: 99%

Inhibitory Effects of Citrus hassaku Extract and Its Flavanone Glycosides on Melanogenesis

Itoh

Hirata

Masuda

et al. 2009

Biological & Pharmaceutical Bulletin

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Melanogenesis stimulated by UV irradiation occurs in plants, microorganisms, and mammalian cells by an enzymatic oxidation process starting with L-tyrosine. Various ingredients for skin-whitening cosmetics are developing to reduce melanogenesis. Tyrosinase catalyzes the oxidation of L-tyrosine to 3,4-dihydroxyphenyl-L-alanine (L-DOPA), followed by the oxidation of L-DOPA to dopaquinone, and oxidative polymerization of several dopaquinone derivatives produces melanin. Thus the tyrosinase inhibitor is one of the candidates for reduction of melanogenesis.1) On the other hand, it has been reported that superoxide dismutase (SOD) is one of the key factors that reduce melanin production caused by UV irradiation.2) Therefore tyrosinase inhibitors with SOD-like activity and/or antioxidant activity may be useful ingredients in the field of skin-whitening cosmetics. During our screening program to find a potential tyrosinase inhibitor from natural resources, we reported several crude drugs, such as Glebnia littoralis F. SCHMIDT, Prunus zippeliana M., Myrica rubra S. et ZUCC., and Arctostaphylos uva-ursi L. SPRENGEL, some of which have been applied to cosmetic beauty preparations. 1,[3][4][5] Recently, the tyrosinase inhibitory activities of the peel of Citrus fruit (Citrus unshiu Markovich) and its flavonoids, such as nobiletin, have been reported. 6,7) As a part of our continuous studies on the biological activities of Citrus species, [8][9][10][11][12] we found that a 50% ethanolic extract (CH-ext) obtained from the unripe fruit of Citrus hassaku HORT ex T. TANAKA, which was collected by thinning out in July, exhibited potent mushroom tyrosinase inhibitory activity. Thinning out the unripe fruit of C. hassaku in July is important for a rich harvest of ripe fruit in December. Thus this study was undertaken to examine whether the unripe fruit of C. hassaku collected in July by thinning can be utilized as a plant resource for skin-whitening cosmetic agents, because, to the best of our knowledge, there is no report on the tyrosinase inhibitory activity of C. hassaku. First, to identify the active component, we carried out activity-guided fractionation of the CH-ext using tyrosinase inhibitory assay. For antioxidant activity, SOD-like and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activities of the CH-ext and its flavanone glycosides were also studied. Second, according to the method of Imokawa, 13) we examined the effects of the CH-ext on melanogenesis using cultured murine B16 melanoma cells after exposure to glucosamine. Third, we examined the in vivo preventive effects of the CH-ext against UVB-induced pigmentation of dorsal skin in brownish guinea pigs. 14) MATERIALS AND METHODSReagents Hesperidin, naringin, and neohesperidin were purchased from Sigma-Aldrich Japan (Tokyo, Japan). Narirutin was isolated from fruit of C. unshiu.10) Other chemical and biochemical reagents were of reagent grade and were purchased from Wako Pure Chemical Industries, Ltd. (Osaka, Japan) and/or Nacalai Tesque, Inc. (Kyoto, Japan)...

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“…A375 cells were seeded and treated as reported above, in order to measure extracellular and intracellular melanin according to the method of Hill et al (23). After 72 h of incubation, the culture medium was removed, centrifuged (700 x g, 10 min) and the supernatant was collected for extracellular melanin quantification.…”

Section: Methodsmentioning

confidence: 99%

Inhibition of Mek 1/2 kinase activity and stimulation of melanogenesis by 5,7-dimethoxycoumarin treatment of melanoma cells

Alesiani

Cicconi²,

Mattei³

et al. 2009

Int J Oncol

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Abstract. In this study, the processes of differentiation and melanogenesis induced by 5,7-dimethoxycoumarin in murine (B16) and human (A375) melanoma cells were investigated.Taking into account the previously demonstrated antiproliferative and differentiation activities of this compound, we examined Ras/Raf/Mek/Erk mitogen-activated protein kinase activity following treatment; inhibition of Mek 1/2 kinase activity and subsequent reduction in Erk 1/2 activation were detected in both cell types. We observed melanogenesis induction associated to an increase in cAMP-response elementbinding protein (CREB) and microphthalmia-associated transcription factor (Mitf) expression, both involved in its regulation. Mitf is fundamental for development, survival and differentiation of melanocyte and melanoma, since it regulates transcription of genes encoding for proteins involved in cell cycle progression or in melanogenesis, such as the enzyme tyrosinase. A significant increase of tyrosinase activity was revealed following treatment in B16 but not in A375 cells, although a strong synthesis of melanin was induced by 5,7-dimethoxycoumarin in both cell lines. The treatment induced protoporphyrine IX accumulation involved in melanogenesis since it promotes stability of cAMP. Finally, the Mek 1/2 inhibitor U0126 significantly potentiated growth inhibition of B16 cells triggered by 5,7-dimethoxycoumarin, suggesting that down-regulation of Raf/Mek/Erk pathway sensitizes melanoma cells to 5,7-dimethoxycoumarin treatment.

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Investigation of the Regulation of Pigmentation in α‐Melanocyte‐Stimulating Hormone Responsive and Unresponsive Cultured B16 Melanoma Cells

Cited by 44 publications

References 19 publications

Melanogenesis Stimulation in Murine B16 Melanoma Cells by Umberiferae Plant Extracts and Their Coumarin Constituents

Melanogenesis Stimulation in Murine B16 Melanoma Cells by Umberiferae Plant Extracts and Their Coumarin Constituents

Inhibitory Effects of Citrus hassaku Extract and Its Flavanone Glycosides on Melanogenesis

Inhibition of Mek 1/2 kinase activity and stimulation of melanogenesis by 5,7-dimethoxycoumarin treatment of melanoma cells

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