基于脑微透析技术的川芎-天麻“升降佐制”药性配伍研究

The project is grounded on the traditional Chinese medicine compatibility principle of "lifting-lowering", based on "lowering-lifting combination" of Ligusticum chuanxiong Hort pill (Ligusticum chuanxiong Hort: Gastrodia elata 1:4) and "lifting-lowering combination" of Gastrodia elata pill (Ligusticum chuanxiong Hort : Gastrodia elata4:1) as the subject of the research study. First reasch the different pharmacology to different syndroms(blood statis syndrom and hyperactivity liver yang syndrom) of migraine and evaluate the effect by vascular endothelial cells in vitro calcium channel blocker function and the penetration of the blood brain barrier (BBB) capacity in vivo; then using brain micro dialysis (MD) technology to create electrically stimulated trigeminal ganglion rat induced migraine animal models. Using Tetramethylpyrazine and gastrodin, simultaneously collect the blood brain dialysate and the tetramethylpyrazine-gastrodin concentration using liquid chromatography-mass spectrometry (LC-MS/MS) method for detection at different time points. Ligusticum chuanxiong Hort-Gastrod elata lifting-lowering compatibility affects the differences in composition of the effect of dynamic distribution between blood brain dialysate simultaneously collected at different time points from Pharmacokinetics perspective. The determination of NO、ET、TXB2、6-Keto-PGF1a content level at different time points observed different balance regulating functions of vascular contraction and relaxation (NO/ET、TXB2/6-Keto-PGF1ａ),and the establishment of concentration-time-effect relationship thus, the analysis illustrates the effect of differences in Ligusticum chuanxiong Hort-Gastrodia elata compatibility from pharmacodynamics (PD).

本课题立足于升降药性配伍理论，以"寓降于升"的大川芎丸（川芎:天麻1:4）和"寓升于降"的天麻丸（川芎:天麻4:1）为研究对象，研究对偏头痛不同证型（血瘀证、肝阳上亢证）的药效学差异，评价其透血脑屏障（BBB）异同；采用脑微透析（MD）技术，以川芎嗪、天麻苷元为指标成分，同步收集血/脑透析液，采用液质联用（LC-MS/MS）方法，检测不同时间点川芎嗪、天麻苷元的浓度，从药代动力学（PK）角度，揭示川芎-天麻升降佐制对效应成分在"血-脑"之间的动态分布差异；同步测定血/脑透析液中NO、ET、TXB2、6-Keto-PGF1ａ含量，观测其对血管缩舒平衡（NO/ET、TXB2/6-Keto-PGF1ａ）的不同调节作用，从药效动力学（PD）角度，阐释川芎-天麻升降佐制经时效应、作用靶点的差异。通过研究，旨在揭示川芎-天麻升降佐制的科学内涵，为临床合理使用升降佐制药对提供依据。

本课题从升降药性配伍角度研究大川芎丸、天麻丸（川芎:天麻分别为1:4、4:1），对血瘀、肝阳上亢型偏头痛药效学差异，评价其透血脑屏障异同；以血-脑微透析技术研究川芎嗪（TMP）、阿魏酸（FA）、天麻苷元（HBA）、天麻素（GAS）不同配比在不同证型偏头痛大鼠血-脑中药动学差异。. 药效学研究结果：（1）对肝阳上亢型偏头痛大鼠药效学：天麻:川芎4:1组与1:1组、1:4组比较，血清NOS活性、血、脑中6-Keto-PGF1α含量降低，与1:4组比较，血中TXB2、脑中NOS含量降低，均具有显著性；（2）对血瘀型偏头痛大鼠药效学：川芎:天麻4:1组与1:1、1:4组比较，血中NOS活性分别降低和升高，脑组织NOS活性，NO、ET含量降低；与1:4组比较，CGRP、TXB2含量升高，6-Keto-PGF1α降低，均具有显著性。. 体外透血脑屏障研究结果：高剂量GAS、HBA明显提高各时间点TMP透过率；各剂量TMP明显提高各时间点HBA透过率及达峰时天麻苷元透过率。. 血-脑微透析药动学结果：（1）对血瘀型偏头痛大鼠药代动力学影响：川芎:天麻4:1组与1:1及单味川芎相比，血、脑中TMP的 T1/2、MRT、Cmax、AUC增加；血、脑中MRT、Cmax、AUC,血中FA的T1/2增加，均具有显著性；（2）对肝阳上亢型偏头痛模型大鼠药代动力学影响：天麻-川芎4:1组与1:1及单味药组相比，大鼠血中GAS的Cmax、AUC减小；脑中AUC、MRT增加；大鼠血中HBA的AUC增加，T1/2、MRT减小，均具有显著性；脑中HBA未检测到。. 本课题完成预期研究任务，基本揭示了川芎-天麻升降药性的科学内涵，为临床用药提供了一定指导作用。