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Vitis amurensis Ruprecht root inhibited α-melanocyte stimulating hormone-induced melanogenesis in B16F10 cells.

Jin KS, Oh YN, Hyun SK, Kwon HJ, Kim BW - Nutr Res Pract (2014)

Bottom Line: Anti-melanogenic activity of VARM was analyzed in α-melanocyte stimulating hormone (MSH)-stimulated B16F10 cells through evaluation of antioxidative activity as well as inhibited tyrosinase activity and melanin contents compared with those of kojic acid and arbutin.Betulinic acid isolated from the CH2Cl2 fraction of VARM significantly attenuated α-MSH-induced melanogenesis in a dose dependent manner, which was stronger than that of arbutin used as a positive control.These results indicate that VARM inhibits oxidative stress, tyrosinase activity, and α-MSH-induced melanogenesis in B16F10 cells, due primarily to the active compound, betulinic acid, in the CH2Cl2 fraction.

View Article: PubMed Central - PubMed

Affiliation: Blue-Bio Industry Regional Innovation Center, Dong-Eui University, 176 Eomgwangno, Busanjin-gu, Busan 614-714, Korea.

ABSTRACT

Background/objectives: The root of Vitis amurensis Ruprecht, a sort of wild-growing grape, has been used in oriental medicine for treatment of skin ailments; however, its dermatological activity is not sufficiently understood. The aim of this study was to investigate tyrosinase inhibitory and anti-melanogenic activities of V. amurensis Ruprecht root methanol extract (VARM) in B16F10 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)-stimulated B16F10 cells through evaluation of antioxidative activity as well as inhibited tyrosinase activity and melanin contents compared with 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 production in 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 CH2Cl2 fraction induced the most potent attenuation of melanogenesis without cytotoxicity and the major compound in the CH2Cl2 fraction was identified as betulinic acid. Betulinic acid isolated from the CH2Cl2 fraction of VARM significantly attenuated α-MSH-induced melanogenesis 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 melanogenesis in B16F10 cells, due primarily to the active compound, betulinic acid, in the CH2Cl2 fraction.

No MeSH data available.


Related in: MedlinePlus

Effect of VARM on B16F10 cell viability (A) and α-MSH induced melanogenesis (B). Arbutin is used as a positive control. Values are expressed as mean ± SD (n = 3). *, ** Significantly different from the vehicle treated control and α-MSH treated control, respectively (P < 0.05).
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Figure 2: Effect of VARM on B16F10 cell viability (A) and α-MSH induced melanogenesis (B). Arbutin is used as a positive control. Values are expressed as mean ± SD (n = 3). *, ** Significantly different from the vehicle treated control and α-MSH treated control, respectively (P < 0.05).

Mentions: The cytotoxicity of VARM for 48 hr was determined using a colorimetric MTS assay in B16F10 cells. Fig. 2A shows the percent cell viability of VARM-treated cells compared with that of vehicle treated cells. No remarkable cytotoxicity was observed in cells treated with VARM under the concentration of 200 µg/mL, which was applied for the following cascade of experiments. As shown in Fig. 2B, melanin content of α-MSH stimulated cells was 2.5 times higher than that of vehicle treated cells, indicating that melanogenesis was successfully induced by α-MSH treatment. On the other hand, α-MSH-stimulated melanin production was significantly (P < 0.5) inhibited by treatment with VARM in a dose-dependent manner (Fig. 2B).


Vitis amurensis Ruprecht root inhibited α-melanocyte stimulating hormone-induced melanogenesis in B16F10 cells.

Jin KS, Oh YN, Hyun SK, Kwon HJ, Kim BW - Nutr Res Pract (2014)

Effect of VARM on B16F10 cell viability (A) and α-MSH induced melanogenesis (B). Arbutin is used as a positive control. Values are expressed as mean ± SD (n = 3). *, ** Significantly different from the vehicle treated control and α-MSH treated control, respectively (P < 0.05).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4198962&req=5

Figure 2: Effect of VARM on B16F10 cell viability (A) and α-MSH induced melanogenesis (B). Arbutin is used as a positive control. Values are expressed as mean ± SD (n = 3). *, ** Significantly different from the vehicle treated control and α-MSH treated control, respectively (P < 0.05).
Mentions: The cytotoxicity of VARM for 48 hr was determined using a colorimetric MTS assay in B16F10 cells. Fig. 2A shows the percent cell viability of VARM-treated cells compared with that of vehicle treated cells. No remarkable cytotoxicity was observed in cells treated with VARM under the concentration of 200 µg/mL, which was applied for the following cascade of experiments. As shown in Fig. 2B, melanin content of α-MSH stimulated cells was 2.5 times higher than that of vehicle treated cells, indicating that melanogenesis was successfully induced by α-MSH treatment. On the other hand, α-MSH-stimulated melanin production was significantly (P < 0.5) inhibited by treatment with VARM in a dose-dependent manner (Fig. 2B).

Bottom Line: Anti-melanogenic activity of VARM was analyzed in α-melanocyte stimulating hormone (MSH)-stimulated B16F10 cells through evaluation of antioxidative activity as well as inhibited tyrosinase activity and melanin contents compared with those of kojic acid and arbutin.Betulinic acid isolated from the CH2Cl2 fraction of VARM significantly attenuated α-MSH-induced melanogenesis in a dose dependent manner, which was stronger than that of arbutin used as a positive control.These results indicate that VARM inhibits oxidative stress, tyrosinase activity, and α-MSH-induced melanogenesis in B16F10 cells, due primarily to the active compound, betulinic acid, in the CH2Cl2 fraction.

View Article: PubMed Central - PubMed

Affiliation: Blue-Bio Industry Regional Innovation Center, Dong-Eui University, 176 Eomgwangno, Busanjin-gu, Busan 614-714, Korea.

ABSTRACT

Background/objectives: The root of Vitis amurensis Ruprecht, a sort of wild-growing grape, has been used in oriental medicine for treatment of skin ailments; however, its dermatological activity is not sufficiently understood. The aim of this study was to investigate tyrosinase inhibitory and anti-melanogenic activities of V. amurensis Ruprecht root methanol extract (VARM) in B16F10 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)-stimulated B16F10 cells through evaluation of antioxidative activity as well as inhibited tyrosinase activity and melanin contents compared with 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 production in 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 CH2Cl2 fraction induced the most potent attenuation of melanogenesis without cytotoxicity and the major compound in the CH2Cl2 fraction was identified as betulinic acid. Betulinic acid isolated from the CH2Cl2 fraction of VARM significantly attenuated α-MSH-induced melanogenesis 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 melanogenesis in B16F10 cells, due primarily to the active compound, betulinic acid, in the CH2Cl2 fraction.

No MeSH data available.


Related in: MedlinePlus