基于中药血清药物化学方法的逍遥散治疗黄褐斑的药效物质基础研究

TCM Serum Pharmacochemistry-Based Analysis of the Effective Components of Xiaoyao San For the Therapy of Chloasma

目的:利用中药血清药物化学的方法,初步确定逍遥散治疗黄褐斑的药效物质基础及其作用机理。方法:通过建立逍遥散经口给药后大鼠血清色谱指纹图,确定大鼠血中移行成分的生药来源,并以血清色谱指纹图为指导,以血中移行成分来源生药为原料,定向富集制备逍遥散提取物;采用体外培养的小鼠B16黑素瘤细胞株,以熊果苷为阳性对照药,用MTT法、多巴氧化法、NaOH裂解法评价逍遥散提取物对细胞增殖、酪氨酸酶活性和黑素合成的影响,探讨逍遥散治疗黄褐斑的药效物质基础及作用机理。结果:逍遥散经口给药后大鼠血中发现了14个入血成分,其中4个为血中原型成分,10个为原型入血或代谢产物,分别来自于柴胡、当归、白芍、甘草、茯苓。定向制备的逍遥散提取物对B16细胞酪氨酸酶活性和黑素合成有明显的抑制作用,作用强度呈浓度依赖性。结论:逍遥散的血中移行成分具有抑制B16细胞酪氨酸酶活性和黑素合成的作用,是逍遥散治疗黄褐斑的药效物质基础。

Using the Serum pharmacochemistry method of Chinese Medicine, the material basis and mechanism of Xiaoyao San for chloasma treatment were studied. The serum fingerprint after oral administration of Xiaoyao San was established. According to the constituents absorbed into blood, an extract of Xiaoyao San was prepared. The effects of cellular proliferation, tyrodinase activity and melanin content of Xiaoyao San extract on the B16 melanoma cell line were measured by the MTT, dopa oxidase assay and NaOH-assay, and compared with arbutin as a positive control. Fourteen compounds absorbed into blood after oral administration of Xiaoyao San were identified, including four original compounds of blood and ten original compounds of crude drugs or metabolites .of Radix Bupleuri, Radix Angelicae Sinensis, Radix Paeoniae Alba, Radix Et Rhizoma Glycyrrhizae, Poria. Tyrosinase activity and melanin synthesis on B16 ceils were significantly inhibited by the extract of Xiaoyao San. The results show that the Xiaoyao San constituents absorbed into blood inhibited tyrosinase activity and melanin synthesis on B16 cells, forming the material basis of Xiaoyao San for the treatment of chloasma.