欧前胡素治疗黄褐斑作用机理研究

Research on Mechanism Function of Imperatorin Treating Chloasma

目的:研究欧前胡素对人黑素瘤细胞(A375细胞)合成黑素的影响,探讨白芷主要成分欧前胡素对黄褐斑治疗作用机理。方法:试验分为空白对照组、雌二醇阳性对照组和欧前胡素浓度为1、10、20、40、60、80μmol·L^(-1)给药组,用上述各组处理A375细胞,用MTT法、Na OH裂解法、多巴氧化法和RT-PCR法检测A375细胞活性、黑素含量、酪氨酸酶(TYR)活性、酪氨酸酶相关蛋白-1、2(TRP-1、TRP-2)、MAPK通路细胞外信号调节酶1、2(ERK1、ERK2)以及氨基末端激酶2(JNK2)的信使RNA表达。结果:阳性对照组对A375活性无影响(P>0.05);欧前胡素浓度在40μmol·L^(-1)以上时,可导致A375细胞大量凋亡(P<0.01);浓度在1~20μmol·L^(-1)时,对A375细胞黑素合成及TYR活性有明显抑制作用(P<0.05或P<0.01),浓度为20μmol·L^(-1)的欧前胡素对A375细胞TYR、TRP-1、TRP-2、ERK1、ERK2及JNK2的mRNA表达具有显著抑制作用(P<0.05或P<0.01)。结论:欧前胡素可通过抑制A375细胞中TYR活性或阻断黑素合成信号的传导,治疗或预防黄褐斑。

Objective： The study is aimed to research the influence of imperatorin on melanoma cell （A375 cells）, and explore mechanism function of the roots of Angelica dahurica for treatment of chloasma. Methods： In experiment, control, estradiol and positive control, and experimental groups （ concentration of imperatorin is 1, 10, 20, 40, 60, 80 μmol·L^-1 ） were divided to operate A375 cells. MTr method, NaOH pyrolysis, dopamine oxidation, and RT-PCR method were applied to test A375 cells activity, melanin content, tyrosine （TYR） enzyme activity, and RNA expression of tyrosinase related protein- 1, 2（ TRP-1, TRP-2 ）, mark pathway extracellular signal regulating enzymes 1, 2 （ERKI, ERK2 ）and Jun N-Terminal Kinase 2 （ JNK2 ）. Results： There existed no significance of A375 cells in positive control group（ P 〉 0. 05 ）. The concentration of imperatorin was above 40 μmol·L^-1 , a lots of A375 cells died（P 〈0. 01 ）. The concentration of imperatorin was 1-20 μmol·L^-1 , there was significant inhibiting effect on A375 cells melanoma cells synthesis and TYR activity. The concentration of imperatorin was 20 μmol·L^-1, there was significant inhibiting effect on TYR, TRP-1, TRP-2, ERK1, ERK2 and mRNA of JNK2 in A375 cells （P 〈 0. 05 or P 〈 0. 01 ）. Conclusion： Imperatorin can treat and prevent chloasma by inhibiting TYR activity in 375 cells or blocking melanin synthesis signal conduction