N-乙酰氨基葡萄糖对酪氨酸酶活性及小鼠B16黑素瘤细胞黑素合成的影响  

Effects of N-acetylglucosamine on tyrosinase activity and melanogenesis in B16 melanoma cells

作　　者：涂彩霞[1] 张荣鑫[1] 刘敏[2] 李国艳[1] 黄畋[1] 

机构地区：[1]大连医科大学附属第二医院皮肤科,116027 [2]大连医科大学附属第二医院中心实验室,116027

出　　处：《中华皮肤科杂志》2009年第9期632-634,共3页Chinese Journal of Dermatology

基　　金：中国宝洁公司资助课题

摘　　要：目的探讨N-乙酰氨基葡萄糖对体外酪氨酸酶活性、小鼠B16黑素瘤细胞酪氨酸酶和黑素合成的影响。方法取对数生长期小鼠B16黑素瘤细胞，分别加入不同浓度N-乙酰氨基葡萄糖（1．5、3、6．25、25、50和100mmol／L）共孵育，MTT法测定培养的小鼠B16黑素瘤细胞的增殖活性，NaOH法测定黑素生成量，体外氧化多巴反应方法测定酪氨酸酶活性；免疫细胞化学测定小鼠B16黑素瘤细胞中酪氨酸酶的表达。结果与阴性对照（0．8183±0．0326）相比，1．5、3、6．25、25和50mmol／LN-乙酰氨基葡萄糖显著抑制蘑菇酪氨酸酶活性（分别为0．7380±0．0254、0．7293±0，0382、0．7247±0．0389、0．7233±0．0186、0．7043±0．0166）（P〈0．05或P〈0．01）；25、50和100mmol／LN-乙酰氨基葡萄糖显著抑制小鼠B16黑素瘤细胞增殖（分别为0．5410±0．0496、0．4480±0．0246、0．1273±0．0137）和黑素生成（0．0070±0．0008、0．0049±0．0012、0．0015±0．0014），50和100mmol／LN-乙酰氨基葡萄糖显著抑制B16黑素瘤细胞酪氨酸酶活性（分别为0．1003±0．0404、0．0130±0．0053）及酪氨酸酶蛋白的表达（13．2700±0．9741、8．5667±2．0345，P〈0．05或P〈0．01）。结论N-乙酰氨基葡萄糖抑制培养的小鼠B16黑素瘤细胞的酪氨酸酶活性和黑素合成。Objective To study the effects of N-acetylglucosamine on the activity of mushroom tyrosinase in vitro as well as on tyrosinase activity and melanogenesis in B16 melanoma cells. Methods Various concentrations （1.5 - 50 mmol/L） of N-acetylglucosamine were used to incubate with mushroom tyrosinase for 10 minutes following by the measurement of dopa oxidase activity of tyrosinase. B16 melanoma cells were cultured and treated with six concentrations （1.5 - 100 mmol/L） of N-acetylglueosamine for 72 hours; then, the cell proliferation of, melanogenesis and expression of tyrosinase protein and activity of tyrosinase in cultured B 16 cells were measured by methyl thiazolyl tetrazolium （MTT） assay, sodium hydroxide （NaOH）-lysis method, immunocytochemical method and dopa oxidation assay, respectively. Results N-acetylglucosamine of 1.5, 3, 6.25, 25 and 50 mmol/L significantly inhibited the activity of mushroom tyrosinase compared with phosphate buffered saline （0.7380 ± 0.0254, 0.7293 ± 0.0382, 0.7247 ± 0.0389, 0.7233 ± 0.0186, 0.7043 ± 0.0166 vs 0.8183 ± 0.0326, P 〈 0.05 or 0.01）. After treatment with N-acetylglucosamine of 25, 50 and 100 mmoFL, a significant suppression was observed in cell proliferation （absorbance： 0.5410 ± 0.0496, 0.4480 ± 0.0246 and 0.1273 ± 0.0137 vs 0.6523 ± 0.0569） of and melanogenesis （absorbance at 460 nm： 0.0070 ± 0.0008, 0.0049 ± 0.0012 and 0.0015 ± 0.0014 vs 0.0096 ± 0.0014） in B16 melanoma cells. Also, decreased activity （absorbance at 460 nm： 0.1003 ± 0.0404 and 0.0130 ± 0.0053 vs 0.2283± 0.0691） and protein expression （13.2700 ± 0.9741 and 8.5667 ± 2.0345 vs 17.4703 ± 2.0583） of tyrosinase were noted in B16 cells treated with N-acetylglucosamine of 50 and 100 mmol/L. Conelusions These studies show that N-acetylglucosamine inhibits tyrosinase activity and melanogenesis in murine B16 melanoma cells. Hence, N-acetylglucosamine may serve as a skin lightening agent in the future.

关 键 词：肿瘤细胞 培养的 一元酚单氧酶 黑素类 N-乙酰氨基葡萄糖 

分 类 号：R739.5[医药卫生—肿瘤]