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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 investigate the effect of bakuchiol on the melanogenesis of A375 cells and its effect on the signal transduction pathway of mitogen-activated protein kinase (MAPK). Methods A375 cells were cultured in vitro and treated with different concentrations of bakuchiol (100μmol / L, 10μmol / L, 1μmol / L, 10-1μmol / L, 10-2μmol / L, -3) μmol / L), the cell proliferation rate and tyrosinase (TYR) activity were detected by MTT method and dopa oxidation method; the change of melanin content was detected by sodium hydroxide cleavage method; The expressions of TRP-1, TRP-2 and MAPK pathway-related protein kinases JNK2, ERK1 and ERK2 were detected by ELISA. Results Compared with the blank group, the melanin biosynthesis rate and TYR activity were significantly higher than those of the blank group (10-1μmol / L, 10-2μmol / L, 10-3μmol / L) The mRNA expression of TYR, TRP-1, TRP-2, JNK2, ERK1 and ERK2 were significantly decreased (P <0.05, P <0.01) and that of bakuchiol (10-1 μmol / L) Inhibition (P <0.05, P <0.01). CONCLUSION: Psoralen can inhibit the melanin synthesis and TYR activity of A375 cells. The changes may be related to down-regulation of mRNA expression of TYR, TRP-1, TRP-2, JNK2, ERK1 and ERK2.