Vitis amurensis Ruprecht root inhibited α-melanocyte stimulating hormone-induced melanogenesis in B16F10 cells
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BACKGROUND/OBJECTIVES: The root of Vitis amurensis Ruprecht, a sort of wild-growing grape, has been used in oriental medicinefor treatment of skin ailments; however, its dermatological activity is not sufficiently understood. The aim of this study wasto investigate tyrosinase inhibitory and anti-melanogenic activities of V. amurensis Ruprecht root methanol extract (VARM) inB16F10 mouse melanoma cells and to attempt to isolate and identify the active compound issued from VARM. MATERIALS/METHODS: Anti-melanogenic activity of VARM was analyzed in α-melanocyte stimulating hormone (MSH)-stimulatedB16F10 cells through evaluation of antioxidative activity as well as inhibited tyrosinase activity and melanin contents comparedwith those of kojic acid and arbutin. After anti-melanogenic analysis of VARM, serial fractionation, nuclear magnetic resonance(NMR), and thin layer chromatorgraphy (TLC) were applied for identification of active compounds contained in VARM. RESULTS: VARM significantly inhibited oxidative stress and tyrosinase activity and attenuated α-MSH-induced melanin productionin B16F10 cells. For isolation of active compounds, VARM was fractionated using a series of organic solvents, including dichloromethane(CH₂Cl₂), ethyl acetate (EtOAc), and n-butanol (n-BuOH). Among fractions showing anti-melanogenic activity, the CH₂Cl₂fractioninduced the most potent attenuation of melanogenesis without cytotoxicity and the major compound in the CH₂Cl₂fractionwas identified as betulinic acid. Betulinic acid isolated from the CH₂Cl₂fraction of VARM significantly attenuated α-MSH-inducedmelanogenesis in a dose dependent manner, which was stronger than that of arbutin used as a positive control. CONCLUSIONS: These results indicate that VARM inhibits oxidative stress, tyrosinase activity, and α-MSH-induced melanogenesisin B16F10 cells, due primarily to the active compound, betulinic acid, in the CH₂Cl₂fraction.
Abstract
BACKGROUND/OBJECTIVES: The root of Vitis amurensis Ruprecht, a sort of wild-growing grape, has been used in oriental medicinefor treatment of skin ailments; however, its dermatological activity is not sufficiently understood. The aim of this study wasto investigate tyrosinase inhibitory and anti-melanogenic activities of V. amurensis Ruprecht root methanol extract (VARM) inB16F10 mouse melanoma cells and to attempt to isolate and identify the active compound issued from VARM. MATERIALS/METHODS: Anti-melanogenic activity of VARM was analyzed in α-melanocyte stimulating hormone (MSH)-stimulatedB16F10 cells through evaluation of antioxidative activity as well as inhibited tyrosinase activity and melanin contents comparedwith those of kojic acid and arbutin. After anti-melanogenic analysis of VARM, serial fractionation, nuclear magnetic resonance(NMR), and thin layer chromatorgraphy (TLC) were applied for identification of active compounds contained in VARM. RESULTS: VARM significantly inhibited oxidative stress and tyrosinase activity and attenuated α-MSH-induced melanin productionin B16F10 cells. For isolation of active compounds, VARM was fractionated using a series of organic solvents, including dichloromethane(CH₂Cl₂), ethyl acetate (EtOAc), and n-butanol (n-BuOH). Among fractions showing anti-melanogenic activity, the CH₂Cl₂fractioninduced the most potent attenuation of melanogenesis without cytotoxicity and the major compound in the CH₂Cl₂fractionwas identified as betulinic acid. Betulinic acid isolated from the CH₂Cl₂fraction of VARM significantly attenuated α-MSH-inducedmelanogenesis in a dose dependent manner, which was stronger than that of arbutin used as a positive control. CONCLUSIONS: These results indicate that VARM inhibits oxidative stress, tyrosinase activity, and α-MSH-induced melanogenesisin B16F10 cells, due primarily to the active compound, betulinic acid, in the CH₂Cl₂fraction.
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