龙眼果核中没食子酸的体外美白作用及机制探索  

In Vitro Whitening Effect and Mechanism of Gallic Acid from Dimocarpus longan Stone

作　　者：黄海潮[1] 郑公铭[1] 张小红[1] 聂阳[1] 巫玮[1] 

机构地区：[1]广东食品药品职业学院,广州510520

出　　处：《中国实验方剂学杂志》2015年第19期121-124,共4页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：广东省科技计划项目(2010B020312006);广东省中医药管理局科研究项目(20121105)

摘　　要：目的：观察从龙眼果核中提取分离的没食子酸对小鼠黑色素瘤B16细胞的作用及对其黑色素合成的影响,探索其作用机制。方法：体外培养小鼠黑色素瘤B16细胞,利用不同质量浓度没食子酸处理一定时间,采用四甲基唑氮蓝（MTT）法观察其对细胞增殖的影响,测定细胞内黑色素的含量变化,观察没食子酸对黑色素合成的影响;通过形态学观察,Hoechst33342染色法及逆转录聚合酶链式反应判断药物抑制细胞的作用方式以及对细胞中酪氨酸酶表达变化的影响,研究其减少黑色素分泌的作用机制。结果：当没食子酸质量浓度1~64 mg·L-1时,药物对B16细胞抑制作用变化明显,药物作用24,48,72 h时的半抑制浓度分别为32.29,22.47,18.68 mg·L-1。没食子酸能明显减少B16细胞中黑色素的含量,在一定范围内,与药物作用的质量浓度及作用时间呈正相关;其对B16细胞的抑制作用及黑色素分泌的影响在不同质量浓度下通过不同方式起作用,在较低质量浓度（≤16 mg·L-1）时,药物通过凋亡的方式起抑制作用,在较高质量浓度（〉16 mg·L-1）时,其通过直接杀伤的方式起抑制作用;在没食子酸质量浓度≤32 mg·L-1时,能明显减少B16细胞中酪氨酸酶的表达。结论：没食子酸对B16细胞具有明显的抑制作用,并可降低细胞中黑色素的含量。没食子酸对B16细胞中酪氨酸酶表达具有明显抑制作用。Objective ： To study effect and mechanism of gallic acid which extracted and separated from Dimocarpus longan stone on melanin synthesis of B16 ceils. Method： Cultured mouse melanoma B16 cells in vitro, and treated with different concentrations of gallic acid in different time. Its effect on B16 cells proliferation was observed by MTT assay, content change of B16 cells melanin was determined to observe effects of gallic acid on melanin synthesis. By morphological observation, Hoechst 33342 staining and reverse transcription polymerase chain reaction （RT-PCR） were adopted to determine inhibition ways of gallic acid on B16 cells, and changes of tyrosinase expression in B16 cells. Mechanism of gallic acid on B16 cells melanin secretion was investigated. Result： Gallic acid had inhibitory effects on B16 cells significantly at the concentration range of 1-64 mg .L-1, half inhibiting concentration （IC50） of gallic acid on B16 cells growth were 32.29, 22.47, 18.68 mg .L-1 after treated for 24, 48, 72 h, respectively. Gallic acid significantly reduced melanin synthesis in B16 cells, and had positively related with concentration and acting time in a certain range. Inhibitory effect of gallic acid on B16 cells and melanin synthesis in different ways with different concentrations. At low concentrations （ ≤ 16 mg .L-1 ）, its inhibitory effect through apoptosis, at high concentration （ 〉 16 mg . L-1）, its inhibitory effect through cell directly killing way. When the concentration of gallic acid ≤ 32 mg .L-1, it significantly reduced expression of tyrosinase in B16 cells. Conclusion： Gallic acid has obvious inhibitory effect on B16 cells and can reduce thecontent of melanin in B16 cells. Gallic acid has obvious inhibitory effect on tyrosinase expression in B16 cells.

关 键 词：没食子酸 龙眼 美白作用 分子机制 细胞凋亡 半抑制浓度 

分 类 号：R285.5[医药卫生—中药学] R945[医药卫生—中医学]