4种美白剂对皮肤黑素细胞的细胞毒作用研究

Cytotoxicity of Four Skin Lighteners on Skin Melanoma Cells

目的探讨4种常用皮肤美白剂(熊果苷、曲酸衍生物、BL-99和氢醌酯)对皮肤黑素细胞的细胞毒作用。方法选用B-16黑素瘤细胞株,分别加入不同剂量(BL-99和曲酸衍生物的实验终浓度分别为0．5、 1．0、5．0、10．0、50．0μg/mL;熊果苷为10、30、50、70、100μg/mL;氢醌酯为0．1、0．5、1．0、5．0、10．0μg/mL)美白剂 3d后,采用台盼蓝排斥法计数活细胞数,采用丙酮酸比色法测定细胞培养上清液中的乳酸脱氢酶(LDH)释放水平。结果 4种美白剂均可使培养细胞数量明显减少,细胞增殖抑制率明显上升。熊果苷、曲酸衍生物和 BL-99等三种美白剂均可导致培养细胞释放LDH水平明显升高,氢醌酯在低浓度(0．1-0．5μg/mL)时引起 B-16细胞LDH释放水平明显高于对照组,而浓度超过1μg/mL时细胞LDH释放水平则呈下降趋势。结论 4种皮肤美白剂均可抑制黑素细胞的生长,具有明显的细胞毒性。在实际使用时建议熊果苷以低于70μg/mL、曲酸衍生物以低于50μg/mL、BL-99以低于10μg/mL剂量为宜,并在美白化妆品和临床用药中不使用或严格限量使用氢醌酯。

Objective To investigate the cytotoxicity of four skin lighteners, such as arbutin, kojic acid derivatives, BL - 99 and hydroquinone dipropionate on murine melenoma cells. Methods Affter B - 16 murine melanoma cells were treated with BL - 99 and kojic acid derivatives in contents of 0.5, 1.0, 5.0, 10.0 and 50.0 μg/mL, arbutin in contents of 10, 30, 50, 70 and 100 μg/mL, hydroquinone dipropionate in contents of 0.1, 0.5, 1.0, 5.0 and 10.0 μg/mL, respectively, for three days, trypan blue exclusion test was used to determine the cell growth, pyruvic acid colorimetry was used to determine the release of cytoplasmic enzyme （Lactate dehydrogenase, LDH）. Results After treated with the four skin lighteners, the total number of cells decreased and the inhibitory rate of cell proliferation increased significantly. The release of LDH increased significantly after treated with arbutin, kojic acid derivatives and BL - 99. Hydroquinone dipropionate at low concentration （0.1 - 0.5μg/mL） led a higher LDH release compared with control group, but that at high concentration （ 1 - 10μg/ mL） the release had a downward trend. Conclusion The four skin lighteners induce cytotoxicity on B - 16 murine melanoma cells and inhibit the melanoma cell＇s proliferation obviously. The suggested dose of arbutin, kojic acid derivatives, and BL- 99 for skin lightening is less than 70 μg/mL, 50 μg/mL , 10 μg/mL, respectively, and hydroquinone dipropionate should not be used for lightening cosmetics and/or drugs, or the dose should be limited strictly.