Arbutin Increases the Pigmentation of Cultured Human Melanocytes Through Mechanisms Other Than the Induction of Tyrosinase Activity

Nakajima, Miki; Shinoda, Ichizo; Fukuwatari, Yasuo; Hayasawa, Hirotoshi

doi:10.1111/j.1600-0749.1998.tb00705.x

Cited by 69 publications

(50 citation statements)

References 19 publications

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“…According to Nakajima et al, 13) arbutin at concentrations in the range of 0.5-8 mM increased the pigmentation of cultured melanocytes. As cosmetics are usually used daily, the effects of the ingredients of cosmetics applied at concentrations in the high-dose range should be examined.…”

Section: Resultsmentioning

confidence: 99%

Inhibitory Effects of Active Compounds Isolated from Safflower (Carthamus tinctorius L.) Seeds for Melanogenesis

Roh¹,

Han²,

Kim

et al. 2004

Biological & Pharmaceutical Bulletin

178

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Section: Resultsmentioning

confidence: 99%

Inhibitory Effects of Active Compounds Isolated from Safflower (Carthamus tinctorius L.) Seeds for Melanogenesis

Roh¹,

Han²,

Kim

et al. 2004

Biological & Pharmaceutical Bulletin

178

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“…Reverse Transcription-Polymerase Chain Reaction (RT-RCR) Total RNA was prepared using a QuickPrep Total RNA Extraction kit (Amersham Pharmacia Biotech) from HMV-II cells grown in the presence or absence of 0.5 mM a-arbutin for 6 d. Two hundred nanograms of the total RNA were reverse-transcribed and the synthesized cDNA was amplified using Ready-to-GO RT-PCR beads (Amersham Pharmacia Biotech) in a total volume of 50 ml containing 0.5 mg pd(T) [12][13][14][15][16][17][18] -first strand primer, 400 nM sense primer and 400 nM antisense primer. Programmable Thermal Cycler conditions were 42°C for 30 min, 95°C for 5 min, 18, 20, 22 or 24 cycles at 95°C for 0.5 min, 60°C for 0.5 min and 72°C for 2 min, and a final treatment at 72°C for 7 min.…”

Section: Assay Of Melanin Synthesis By Hmv-ii Cellsmentioning

confidence: 99%

Inhibitory Effects of .ALPHA.-Arbutin on Melanin Synthesis in Cultured Human Melanoma Cells and a Three-Dimensional Human Skin Model

Sugimoto

Nishimura

Nomura

et al. 2004

Biological & Pharmaceutical Bulletin

130

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Hyperpigmentation in the epidermis is caused by excessive melanin synthesis, and tyrosinase (EC 1.14.18.1) is a key-enzyme in melanin synthesis. It catalyzes the first two steps in melanin synthesis: the hydroxylation of tyrosine to 3-(3,4-dihydroxyphenyl)-alanine (DOPA) and the oxidation of DOPA to dopaquinone. Several tyrosinase inhibitors have been used as skin-lightening agents in the cosmetic industry.4-Hydroxyphenyl a-D-glucopyranoside (a-arbutin; Fig. 1) was enzymatically synthesized from hydroquinone and saccharides, [1][2][3][4] and its inhibitory activity against tyrosinases from mushroom, B16 mouse melanoma and HMV-II human melanoma cells has been examined previously.2,5,6) a-Arbutin specifically inhibited mammalian tyrosinases, and its effect on human tyrosinase was the strongest among the hydroquinone-glycosides that we have studied so far. 6) In this study, we examined the inhibitory effects of a-arbutin on melanin biosynthesis in cultured human melanoma cells and in a three-dimensional human skin model. MATERIALS AND METHODSCell Culture Human malignant melanoma cells, HMV-II 7) were provided by the Cell Resource Center for Biomedical Research, Tohoku University. The cells were cultured in F12/DMEM (GIBCO) supplemented with 10% fetal bovine serum at 37°C in a humidified atmosphere of 5% CO 2 in air.The three-dimensional cultured human skin model, MelanoDerm, (MEL312B; a co-culture of normal human melanocytes (NHM) from black donors and normal human keratinocytes (NHK)) was purchased from Kurabo Co. (Osaka, Japan). The human skin model was maintained according to the manufacturer's instructions.Reagents L-DOPA and synthetic melanin were purchased from Sigma Chemical Co. (St. Louis, MO, U.S.A.). Medium for the human skin model [long life maintenance medium (LLMM)] was supplied with the MEL-312B kit. Other chemicals were purchased from Wako Pure Chemical Industries, Ltd. (Osaka).Preparation of a a-Arbutin a-Arbutin was prepared by the method described by Nishimura et al.1) a-Amylase from Bacillus sp. strain X-23 was added to 100 ml of 50 mM sodium acetate buffer solution (pH 5.0) containing 5% hydroquinone and 20% maltopentaose. After incubation at 40°C for 16 h, glucoamylase from Aspergillus niger was added, and the mixture was incubated at 40°C for 4 h. a-Arbutin was purified from the reaction mixture by extraction with ethyl acetate and charcoal column chromatography.Assay of Melanin Synthesis by HMV-II Cells HMV-II cells (1.0ϫ10 6 ) were plated with 10 ml of medium in a 100-mm dish and grown for 10 d. On days 1, 4 and 7, the medium was changed to fresh medium containing various concentrations of a-arbutin.Determination of Melanin Content The melanin content of cells was measured by the method of Hosoi et al. -free phosphate-buffered saline [PBS(Ϫ)]. After being treated with 0.25% trypsin, the cells were harvested by centrifugation at 1000ϫg for 10 min and sonicated in 0.5 ml of 1% Triton X-100/PBS(Ϫ) on ice. In contrast, the human skin model was harvested and sonicated in 0.5 ml of PBS(Ϫ). The lys...

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“…Cell lysates were analyzed for tyrosinase activity through assay of a chromogenic substrate, as described by Nakajima (19), measured by absorption spectrophotometry at 490 nm.…”

Section: Tyrosinase Activity Assaymentioning

confidence: 99%

Regulation of melanocyte apoptosis by Stathmin 1 expression

Zhang

Xiong²,

Wang³

et al. 2008

BMB Reports

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Arbutin Increases the Pigmentation of Cultured Human Melanocytes Through Mechanisms Other Than the Induction of Tyrosinase Activity

Cited by 69 publications

References 19 publications

Inhibitory Effects of Active Compounds Isolated from Safflower (Carthamus tinctorius L.) Seeds for Melanogenesis

Inhibitory Effects of Active Compounds Isolated from Safflower (Carthamus tinctorius L.) Seeds for Melanogenesis

Inhibitory Effects of .ALPHA.-Arbutin on Melanin Synthesis in Cultured Human Melanoma Cells and a Three-Dimensional Human Skin Model

Regulation of melanocyte apoptosis by Stathmin 1 expression

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