驱虫斑鸠菊黄酮促人表皮黑素细胞黑素合成及其机制的研究

The effect of vernonia anthelmintica flavone on melanogenesis of human melanocytes in vitro

目的:观察驱虫斑鸠菊黄酮对体外培养的人表皮黑素细胞(MC)黑素合成和酪氨酸酶活性的影响,并初步探讨其可能的作用机制。方法:噻唑兰比色法(MTT)测定药物对细胞增殖率的影响,选择高、中、低3种浓度(30、10、3μg/ml)的驱虫斑鸠菊黄酮作用体外培养的人表皮黑素细胞,观察对黑素细胞的形态、酪氨酸酶活性、黑素含量的影响。免疫印迹法测定药物作用前后MC酪氨酸酶(TYR)、酪氨酸酶相关蛋白1(TRP1)和酪氨酸酶相关蛋白2(TRP2)蛋白表达的变化。结果:≥90μg/ml驱虫斑鸠菊黄酮可明显抑制MC的增殖;试验浓度范围内可明显促进MC黑素合成和酪氨酸酶活性增加,并以浓度依赖方式促进TRP-1的蛋白表达,但对TYR、TRP-2的表达没有影响。结论:驱虫斑鸠菊黄酮在体外能直接刺激表皮MC黑素合成,上述变化可能通过促进TRP-1的蛋白表达和TYR的翻译后修饰作用来完成。

Objective To investigate the effect of vernonia anthelmintica flavone （VAF） on melanogenesis and tyrosinase activity of human epidermal melanocytes, and initially discuss their related mechanism. Methods Cell proliferation was observed by MTT method. Three different concentrations of VAF were incubated with human melanocytes for 72h and then melanin content, tyrosinase activity were respectively measured. The protein expression of melanogenesis key enzymes was investigated by western blot. Results The proliferation of melanocytes was most inhibited by VAF at high dose （≥1001μg/ml）. In the range of experimental dose, VAF increased the activity of the tyrosinase and melanin content significantly in a dose-dependent manner. The increasing protein expression of TRP1 was also observed in a dose-dependent manner. No effect was observed on protein expression of TYR and TRP2. Conclusion VAF may stimulate melanogenesis directly on human epidermal melanocytes, which may be induced by increasing expression of TPR1 and post-translational modification of TYR.