补骨脂酚对人A375细胞黑素生成及MAPK信号通路干预作用研究

The research of bakuchiol on melanin synthesis and its intervention of MAPK signaling pathway in human A375 cells

目的探讨补骨脂酚抑制A375细胞的黑素合成效果及对丝裂原活化蛋白激酶(MAPK)信号转导通路的干预作用。方法体外培养A375细胞,经不同浓度的补骨脂酚(100μmol/L、10μmol/L、1μmol/L、10^(-1)μmol/L、10^(-2)μmol/L、10^(-3)μmol/L)干预后,采用MTT法和多巴氧化法检测细胞增殖率和酪氨酸酶(TYR)活性;氢氧化钠裂解法检测黑素含量的变化;RT-PCR法测定酪氨酸酶及相关蛋白(TRP-1、TRP-2)和MAPK通路相关蛋白激酶JNK2、ERK1、ERK2 mRNA的表达量。结果与空白组比较,补骨脂酚(10^(-1)μmol/L、10^(-2)μmol/L、10^(-3)μmol/L)对黑素合成率和TYR活性均明显降低(P<0.05,P<0.01),补骨脂酚(10^(-1)μmol/L)对TYR、TRP-1、TRP-2、JNK2、ERK1、ERK2的mRNA基因表达量均显著抑制(P<0.05,P<0.01)。结论补骨脂酚对A375细胞黑素合成及TYR活性有抑制作用,其变化可能是通过下调TYR、TRP-1、TRP-2、JNK2、ERK1、ERK2的mRNA基因表达量来实现的。

Objective To discuss bakuchiol on melanin synthesis and its intervention of MAPK signaling transduction pathways in A375 cells. Methods The cultured A375 cells were treated with bakuchiol; MTT assay,L-dopa oxidation assay were respectively used to detect the activity of cell and TYR; odium hydroxide lysis assay were used to detect the change of melanin synthesis; RT-PCR was applied to measure the mRNA expression levels of TYR,TRP-1,TRP-2 and the mRNA expression levels of JNK2,ERK1,ERK2. Results bakuchiol（ 10- 1μmol /L,10- 2μmol /L,10-3μmol /L） can inhibit melanin synthesis and the activity of TYR,Bakuchiol（ 10- 1μmol /L） can significantly reduce the mRNA expression level of TYR,TRP-1,TRP-2,JNK2,ERK1 and ERK2（ P 〈 0. 05,P 〈 0. 01）.Conclusion Bakuchiol can reduce melanin synthesis and the activity of TYR,and the changes realized probably through reducing the mRNA expression level of TYR,TRP-1,TRP-2,JNK2,ERK1 and ERK2.