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The mechanism of melanocytes-specific cytotoxicity induced by phenol compounds having a prooxidant effect, relating to the appearance of leukoderma.

Nagata T, Ito S, Itoga K, Kanazawa H, Masaki H - Biomed Res Int (2015)

Bottom Line: Furthermore, although raspberry ketone (RK), RD derivative, also increased intracellular ROS in B16F10 cells, increase in ROS was suppressed by disodium dihydrogen ethylenediaminetetraacetate dehydrate (EDTA).The amounts of increased ROS with RK in HaCaT cells without melanocyte were further increased by tyrosinase.Therefore, tyrosinase, a metalloprotein having copper, was speculated to be one of causative agents allowing phenol compounds to work as a prooxidant.

View Article: PubMed Central - PubMed

Affiliation: Institute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University, 2-10 Kawadacho, Shinjuku-ku, Tokyo 162-0054, Japan ; I.T.O. Co. Ltd., 1-6-7-3F Naka-cho, Musashino, Tokyo 180-0006, Japan.

ABSTRACT
Specific phenol compounds including rhododendrol (RD), a skin-brightening ingredient in cosmetics, are reported to induce leukoderma, inducing a social problem, and the elucidation of mechanism of leukoderma is strongly demanded. This study investigated the relationship among the cytotoxicities of six phenol compounds on B16F10 melanoma cells and HaCaT keratinocytes and generated reactive oxygen species (ROS). As a result, the cytotoxicity of RD on B16F10 cells was higher than that on HaCaT cells, and RD significantly increased intracellular ROS and hydrogen peroxide (H2O2) levels in B16F10 cells. Furthermore, although raspberry ketone (RK), RD derivative, also increased intracellular ROS in B16F10 cells, increase in ROS was suppressed by disodium dihydrogen ethylenediaminetetraacetate dehydrate (EDTA). The amounts of increased ROS with RK in HaCaT cells without melanocyte were further increased by tyrosinase. Therefore, tyrosinase, a metalloprotein having copper, was speculated to be one of causative agents allowing phenol compounds to work as a prooxidant. Hydroxyl radical was generated by adding a mixture of tyrosinase and H2O2 to RD, and the amount of the radical was further increased by UVB, indicating that RD cytotoxicity was caused by intracellularly increased ROS, which possibly related to phenol induced prooxidants.

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Related in: MedlinePlus

Effect of tyrosinase on intracellular reactive oxygen species (ROS) induced by raspberry ketone (RK). After HaCaT keratinocytes were treated with 50.0~100.0 units/mL tyrosinase, 1.0 mM RK, and 20.0 μM 2′,7′-dichlorofluorescin diacetate (DCFDA) for 30 min, the fluoresce intensities were measured. The black and gray columns indicate the averaged intracellular ROS levels in B16F10 treated with 0 and 1.0 mM RK, respectively. The lines on the columns show the standard deviations (n = 5). Two asterisks (∗∗) indicate that the probabilities of significant levels are less than 0.01 (P < 0.01).
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fig6: Effect of tyrosinase on intracellular reactive oxygen species (ROS) induced by raspberry ketone (RK). After HaCaT keratinocytes were treated with 50.0~100.0 units/mL tyrosinase, 1.0 mM RK, and 20.0 μM 2′,7′-dichlorofluorescin diacetate (DCFDA) for 30 min, the fluoresce intensities were measured. The black and gray columns indicate the averaged intracellular ROS levels in B16F10 treated with 0 and 1.0 mM RK, respectively. The lines on the columns show the standard deviations (n = 5). Two asterisks (∗∗) indicate that the probabilities of significant levels are less than 0.01 (P < 0.01).

Mentions: Tyrosinase was investigated to be involved in the increase in intracellular ROS level. As a result, 50.0, 75.0, and 100.0 units/mL tyrosinase were confirmed to further promote increase in the intracellular ROS level in HaCaT cells treated with 1.0 mM RK (Figure 6).


The mechanism of melanocytes-specific cytotoxicity induced by phenol compounds having a prooxidant effect, relating to the appearance of leukoderma.

Nagata T, Ito S, Itoga K, Kanazawa H, Masaki H - Biomed Res Int (2015)

Effect of tyrosinase on intracellular reactive oxygen species (ROS) induced by raspberry ketone (RK). After HaCaT keratinocytes were treated with 50.0~100.0 units/mL tyrosinase, 1.0 mM RK, and 20.0 μM 2′,7′-dichlorofluorescin diacetate (DCFDA) for 30 min, the fluoresce intensities were measured. The black and gray columns indicate the averaged intracellular ROS levels in B16F10 treated with 0 and 1.0 mM RK, respectively. The lines on the columns show the standard deviations (n = 5). Two asterisks (∗∗) indicate that the probabilities of significant levels are less than 0.01 (P < 0.01).
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4377363&req=5

fig6: Effect of tyrosinase on intracellular reactive oxygen species (ROS) induced by raspberry ketone (RK). After HaCaT keratinocytes were treated with 50.0~100.0 units/mL tyrosinase, 1.0 mM RK, and 20.0 μM 2′,7′-dichlorofluorescin diacetate (DCFDA) for 30 min, the fluoresce intensities were measured. The black and gray columns indicate the averaged intracellular ROS levels in B16F10 treated with 0 and 1.0 mM RK, respectively. The lines on the columns show the standard deviations (n = 5). Two asterisks (∗∗) indicate that the probabilities of significant levels are less than 0.01 (P < 0.01).
Mentions: Tyrosinase was investigated to be involved in the increase in intracellular ROS level. As a result, 50.0, 75.0, and 100.0 units/mL tyrosinase were confirmed to further promote increase in the intracellular ROS level in HaCaT cells treated with 1.0 mM RK (Figure 6).

Bottom Line: Furthermore, although raspberry ketone (RK), RD derivative, also increased intracellular ROS in B16F10 cells, increase in ROS was suppressed by disodium dihydrogen ethylenediaminetetraacetate dehydrate (EDTA).The amounts of increased ROS with RK in HaCaT cells without melanocyte were further increased by tyrosinase.Therefore, tyrosinase, a metalloprotein having copper, was speculated to be one of causative agents allowing phenol compounds to work as a prooxidant.

View Article: PubMed Central - PubMed

Affiliation: Institute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University, 2-10 Kawadacho, Shinjuku-ku, Tokyo 162-0054, Japan ; I.T.O. Co. Ltd., 1-6-7-3F Naka-cho, Musashino, Tokyo 180-0006, Japan.

ABSTRACT
Specific phenol compounds including rhododendrol (RD), a skin-brightening ingredient in cosmetics, are reported to induce leukoderma, inducing a social problem, and the elucidation of mechanism of leukoderma is strongly demanded. This study investigated the relationship among the cytotoxicities of six phenol compounds on B16F10 melanoma cells and HaCaT keratinocytes and generated reactive oxygen species (ROS). As a result, the cytotoxicity of RD on B16F10 cells was higher than that on HaCaT cells, and RD significantly increased intracellular ROS and hydrogen peroxide (H2O2) levels in B16F10 cells. Furthermore, although raspberry ketone (RK), RD derivative, also increased intracellular ROS in B16F10 cells, increase in ROS was suppressed by disodium dihydrogen ethylenediaminetetraacetate dehydrate (EDTA). The amounts of increased ROS with RK in HaCaT cells without melanocyte were further increased by tyrosinase. Therefore, tyrosinase, a metalloprotein having copper, was speculated to be one of causative agents allowing phenol compounds to work as a prooxidant. Hydroxyl radical was generated by adding a mixture of tyrosinase and H2O2 to RD, and the amount of the radical was further increased by UVB, indicating that RD cytotoxicity was caused by intracellularly increased ROS, which possibly related to phenol induced prooxidants.

Show MeSH
Related in: MedlinePlus