葡萄籽原花青素抑制小鼠B16-F0黑色素瘤细胞黑色素生成的影响  

The inhibitory effects of grape seed proanthocyanidins on melanogenesis in mouse melanoma B16 cell

作　　者：张慧瑛[1] 邢芙玲 罗光宏[3] 叶生宝 张泽星[1] 邓世川 郝军元[1] 

机构地区：[1]河西学院农业与生物技术学院,张掖734000 [2]张掖市妇幼保健院,张掖734000 [3]河西学院凯源生物技术开发中心,张掖734000 [4]中南林业科技大学生命科学与技术学院,长沙410004

出　　处：《中药药理与临床》2017年第1期63-66,共4页Pharmacology and Clinics of Chinese Materia Medica

基　　金：甘肃省青年科技研究基金项目(1107RJYG012);甘肃省科技支撑计划项目(1304FKCG103);甘肃省科技支撑计划项目(1104FKCG118);甘肃省高校河西走廊特色资源利用省级重点实验室项目(XZ1014)

摘　　要：目的:研究葡萄籽原花青素对小鼠B16-F0黑色素瘤细胞黑色素生成的抑制作用,并对其机理进行初步探究。方法:采用四甲基偶氮唑蓝(MTT)检测小鼠B16黑色素瘤细胞增值率;左旋多巴氧化法测定酪氨酸酶活;Na OH裂解法测定黑色素合成;分别采用RT-PCR法和Western-blotting法检测B16-F0细胞中TYR、TRP-1、TRP-2mRNA和蛋白表达水平。结果:葡萄籽原花青素显著抑制B16黑色素瘤细胞的增殖,且与浓度和作用时间相关;12.5~200μg/ml浓度范围的葡萄籽原花青素显著抑制细胞酪氨酸酶活性及黑色素生成。当葡萄籽原花青素浓度为200μg/ml作用时间为48 h时,对B16-F0细胞内酪氨酸酶活性及黑色素合成抑制效果最佳。此外,与对照组相比,经浓度为100μg/ml和200μg/ml,作用时间为48 h的葡萄籽原花青素处理,B16-F0细胞内TYR、TRP-1、TRP-2的mRNA和蛋白表达水平显著降低。结论:葡萄籽原花青素抑制B16黑色素瘤细胞的增殖及黑色素的合成,其机制可能是通过抑制TYR、TRP-1、TRP-2的表达,进而抑制酪氨酸酶活性实现的。Objective： To investigate the inhibitory effects of grape seed proanthocyanidins on the proliferation,the tyrosinase activity,synthesis of melanin in mouse melanoma B16 cell.Methods： The cell proliferation was determined by MTT assay,and the tyrosinase activity and melanin synthesis were detected by L-dopa oxidation and Na OH assay,respectively.The mRNA and protein expression of tyrosine（ TYR）,Tyrosine associated protease-1（ TRP-1） and Tyrosine associated protease-2（ TRP-2） in B16 cell were analyzed by the reverse transcription-polymerase chain reaction（ RT-PCR） and western-blotting.Results： The cell proliferation was significantly inhibited after treated with grape seed proanthocyanidins（ p 〈 0.05）.The tyrosinase activity and melanin synthesis in B16 cell were significantly inhibited when the concentration of grape seed proanthocyanidins were 12.5 ~ 200 μg/ml.The tyrosinase activity and melanin synthesis were inhibited adequately by the grape seed proanthocyanidins（ 200 μg/ml,48 h）.In addition,the expression of mRNA and protein were significantly down-regulated after treatment with 200μg/ml grape seed proanthocyanidins（ p 〈 0.05）.Conclusion： The grape seed proanthocyanidins could suppress the melanogenesis in B16 cell and the effect may be related with the down-regulation of mRNA and protein expression of TYR,TRP-1,and TRP-2.

关 键 词：葡萄籽原花青素 酪氨酸酶 黑色素生成 酪氨酸酶相关蛋白-1 酪氨酸酶相关蛋白-2 

分 类 号：R285[医药卫生—中药学]