基于定量谱效关系和PK-PD的白芷治疗偏头痛作用机制研究

To deal with the current problems in correlation research between the material basis of pharmacodynamics and the mechanism of action, the genuine regional - Angelica dahurica(Fisch.ex Hoffm.)Benth.et Hook.f.var.formosana (Boiss.) Shan et Yuan - a genuine staple herb grown in Sichuan, will be taken as the object of study based on preliminary studies of this subject. The characteristics of multi-ingredient and multi-target will be fully taken into consideration. The HPLC-MS and other analysis methods will be used together with the virtual screening method and model animal pharmacological activity experiment. The pharmacodynamic ingredients of migraine treatment by Angelica dahurica(Fisch.ex Hoffm.)Benth.et Hook.f.var.formosana (Boiss.) Shan et Yuan will be found out. The functional relationship between pharmacological activity (Y) and the contents of various pharmacodynamic ingredients (Xi) will be established through mathematical method: Y=f(Xi), namely, "quantitative spectrum efficiency relationship". This function will be used to illustrate the complicated relationships between pharmacodynamic ingredients and pharmacological activity. Meanwhile, the changes of pharmacodynamic indicators and the pharmacokinetics characteristics of effective ingredient group will be dynamically monitored after the model animal is intervened by Angelica dahurica(Fisch.ex Hoffm.)Benth.et Hook.f.var.formosana (Boiss.) Shan et Yuan. The model of pharmacokinetics (PK) and pharmacodynamics (PD) will be established to find out the complicated relationships between various effective ingredients of Angelica dahurica(Fisch.ex Hoffm.)Benth.et Hook.f.var.formosana (Boiss.) Shan et Yuan and pharmacodynamic indicators and to reveal the mechanism of migraine treatment by Angelica dahurica(Fisch.ex Hoffm.)Benth.et Hook.f.var.formosana (Boiss.) Shan et Yuan.

针对目前药效物质基础和作用机制相关性研究存在的问题，在前期研究基础上，充分考虑到中药多组分、多靶点的作用特点，以临床疗效确切、有效成分基本明确的川产道地药材白芷为研究对象，采用HPLC-MS等分析手段，结合虚拟筛选方法和模型动物药理活性实验，明确白芷治疗偏头痛的药效成分；通过数学方法建立药理活性（Y）与各药效成分含量（Xi）之间的函数关系：Y=f(Xi)，- - 即"定量谱效关系"，阐明药效成分与药理活性间的复杂关系；同时动态监测白芷干预模型动物后药效指标的变化和有效成分群的药动学特征，构建药物代谢动力学（PK）-药物效应动力学（PD）关系模型，以期明确白芷多个有效成分和药效指标间的复杂对应关系，从而揭示白芷治疗偏头痛作用机制。

在前期研究基础上，以川产道地药材白芷为研究对象，采用HPLC-MS、GC-MS等分析手段，结合虚拟筛选方法和模型动物药理活性实验，建立药理活性与药效成分含量之间的谱效关系；应用实时荧光定量聚合酶链式反应(qPCR)方法对硝酸甘油诱导的偏头痛大鼠中eNOS、c-jun、c-fos、5-DH1D、TNF-a等多种基因表达进行定量分析；对有效成分进行硝酸甘油型偏头痛模型大鼠PK-PD研究，构建药物代谢动力学（PK）-药物效应动力学（PD）关系模型，阐释白芷治疗偏头痛有效成分和多种药效指标间的复杂对应关系。有效成分虚拟筛选和硝酸甘油型偏头痛动物模型验证实验结果显示：欧前胡素、白当归素、东莨菪苷、水合氧化前胡素对硝酸甘油型偏头痛模型有明显影响；基因表达定量分析表明欧前胡素高剂量组能够明显降低NF-κB、CGRP、TNF-a、iNOS、eNOS、c-fos、c-jun基因的mRNA的相对表达量，其余成分对表达量降低作用不明显；PK-PD研究表明，白芷有效成分对造模后外周循环中NO的最终量有显著的降低作用，对造模后脑组织中CGRP的最终量有较弱的降低作用。经研究证实白芷对抗硝酸甘油引起的偏头痛的主要作用途径为抑制外周NO的浓度而产生的，白芷治疗偏头痛的主要作用机制与其降低外周扩血管物质浓度密切相关。本研究从谱效学、基因表达、PK－PD多角度联合对白芷治疗偏头痛作用机制进行研究，从而为进一步的中药作用机制研究提供思路参考。