小柴胡汤对B16黑素瘤细胞黑素合成相关蛋白表达的影响

Effects of Xiaochaihu Decoction on melanogenesis-related proteins of B16 melanoma cells

目的:研究不同浓度小柴胡汤对B16黑素瘤细胞黑素合成相关酶酪氨酸酶(TYR),酪氨酸酶相关蛋白-1(TRP-1),酪氨酸酶相关蛋白-2(TRP-2)蛋白表达的影响。方法:采用CCK8检测细胞活力,选取小柴胡汤低、中、高浓度分别为0.1、1.0、2.5mg/mL用于实验,Western Blot法检测B16细胞中TYR、TRP-1、TRP-2蛋白的表达。结果:与对照组比较,随着浓度的增加,小柴胡汤水煎液能够显著下调细胞TYR、TRP-2蛋白的表达(P<0.05,P<0.01);小柴胡汤中、高浓度组较对照组降低TRP-1活性(P<0.01)。结论:小柴胡汤对黑素合成的抑制作用除了通过直接抑制TYR活性外,还可通过下调酪氨酸酶家族其他成员TRP-1、TRP-2的基因表达、蛋白合成三方面实现其抑制黑素生成的作用,提示小柴胡汤对黑素生成的抑制可通过对酶蛋白转录后调节途径来实现。

Objective:To observe the effects of Xiaochaihu Decoction on melanogenesis-related proteins including tyrosinase(TYR),tyrosinase related proteins-1(TRP-1),tyrosinase related proteins-2(TRP-2) in B16 melanoma cells.Methods:CCK8 assay was used to test the cell activity.The B16 melanoma cells were treated with the administration of Xiaochaihu Decoction at low(0.1mg/mL),medium(1mg/mL) and high(2.5mg/mL) dose,respectively.The protein expressions of TYR,TRP-1,TRP-2 in B16 melanoma cells were detected with Western Blot.Results:Compared with the control group,Xiaochaihu Decoction significantly decreased protein expression levels of TYR,TRP-2 with the increase of its concentration(P<0.05,P<0.01).The protein expression level of TRP-1 in the medium and high does Xiaochaihu Decoction groups were decreased(P<0.01).Conclusion:Besides the direct inhibitive effects on TYR activity,Xiaochaihu Decoction played important roles in the down-regulating the gene and protein expressions of other members in tyrosinase family such as TRP-1 and TRP-2,resulting in the inhibition of melanogenesis.That suggested that the inhibition of melanogenesisby Xiaochaihu Decoction could be realized by regulating the enzyme protein post-transcriptional pathway.