逍遥散中当归的抗抑郁活性成分的研究

Antidepression constituents from Angelica Sinensis Radix in Xiaoyao Powder

目的指认逍遥散中君药当归对复方抗抑郁有效部位的贡献成分,并筛选其中潜在的抗抑郁活性成分。方法采用UPLC-PDA技术对逍遥散抗抑郁有效部位及当归相应部位的指纹图谱进行匹配,通过对比色谱峰的保留时间及全波长紫外光谱图,归属当归对逍遥散抗抑郁有效部位的贡献成分。运用多种色谱方法对贡献成分进行导向分离,采用现代谱学技术鉴定化合物的结构,并对分离得到的化合物进行体外活性验证。结果共归属并鉴定出当归对逍遥散抗抑郁有效部位的贡献成分5个,分别为阿魏酸松柏醇酯(1)、E-丁烯基苯酞(2)、藁本内酯(3)、Z-丁烯基苯酞(4)、12-异戊烯酰基-14-乙酰基-2E,8E,10E-三烯-4,6-二炔-1-醇(5),其中化合物5为首次从伞形科植物中分离得到。化合物1、3、4能有效保护谷氨酸损伤的SH-SY5Y和PC12细胞。结论 UPLC-PDA导向分离技术可快速、准确指认复方来源于其单味药材的贡献成分。阿魏酸松柏酯、藁本内酯、Z-丁烯基苯酞可能为当归贡献逍遥散抗抑郁作用的活性成分。

Objective This current study focused on the identification of active constituents from Angelica Sinensis Radix in Xiaoyao Powder based on UPLC-PDA-guided isolation technique. Methods The UPLC-PDA chromatogram of Xiaoyao Powder was compared with that of Angelica Sinensis Radix. The relative retention time of each peak and the Uhraviolet spectra provided by PDA were used in the analyses. The constituents were isolated from Angelica Sinensis Radix under the guidance of UPLC-PDA investigation. The structures of the isolates were elucidated by NMR techniques. The antidepression effect was evaluated on glutamate-induced neurons. Results Five marker peaks of Xiaoyao Powder fingerprint belonged to Angelica Sinensis Radix and they were determined as coniferyl ferulate（1）, E-butylidenephthalide（2）, ligustilide（3）, Z-butylidenephthalide（4）, and 14-acetoxy-12-senecioyloxytetradeca-2E,8E,10E-trien-4,6-diyn-1-ol（5）. Compound 5 was isolated from the plants in Umbelliferae for the first time. The treatment with compounds 1, 3, and 4 could protect PC12 and SH-SY5 Y cells from glutamate-induced cytotoxicity. Antidepression bioactivity of compound 1 was first investigated. Conclusion UPLC-PDA-guided isolation technique is confirmed to be a rapid and accurate method to identify the main active constituents from Angelica Sinensis Radix in Xiaoyao Powder.