基于血脑PK-PD和结构方程模型的银杏叶提取物多组分协同抗脑缺血作用研究

The synergetic effect of multi-components is the key of traditional Chinese medicine (TCM) to play the overall effect, but the mechanism is needed to be studied further. Our previous study shows that interactions among multi-components can have influence on pharmacokinetic process. So we suppose that changing pharmacokinetic process may be one of the mechanisms of their synergetic effect. How to resolve the complex relations between multi-components’ behavior in vivo and their synergetic effect quantitatively is one of the difficulties in TCM study. In this project, we will select pharmacological effects closely related to cerebral ischemic injury from such levels as whole animal, cell, molecule to evaluate the synergetic effect of ginkgo flavonoids and ginkgo terpene lactones in Ginkgo biloba extract (EGB) on cerebral ischemic injury. Determine pharmacokinetic (PK) and pharmacodynamic (PD) parameters both in blood and brain dialysate. Analyze the influence of ginkgo flavonoids combined with ginkgo terpene (EGB) on multi-components’ absorption and distribution to brain by PK parameters. Use structural equation model (SEM) to resolve causality of multi-components both in blood and brain and the effect quantitatively on basis of PK-PD. Illuminate the protective effect mechanism of multi-components’ synergetic effect in EGB on cerebral ischemic injury. In this project, mechanisms of multi-components’ synergetic effect will be explored from 3 aspects: pharmacological effects, pharmacokinetic process, quantitative relations between multi-components and pharmacological effects, using such methods as multi-components’ PK-PD both in blood and brain, quantitative analysis of mathematical model and etc. It can provide basis for quality control of EGB crude drugs and related preparations. It can also provide references for study of multi-components in TCM.

多组分协同作用是中药发挥整体药效的关键，但其协同作用机制有待探究。前期研究发现，多组分相互作用可影响体内动力学过程，推测体内过程改变是其协同作用机制之一。如何定量解析多组分体内行为和药效间的关系，是中药研究的难点之一。本项目拟从整体动物、细胞、分子层面，选择与脑缺血性损伤密切相关的药效指标，评价银杏叶提取物中黄酮和萜内酯抗脑缺血的协同作用；通过测定血和脑透析液中多组分PK、PD，由PK分析黄酮和萜内酯合用(EGB)对多组分体内过程（吸收、脑内分布）的影响；PK-PD采用结构方程模型定量解析，探索吸收入血和分布入脑的多组分与药效的因果关系，阐明EGB多组分协同保护脑缺血性损伤的作用机制。该项目采用血、脑多组分PK-PD、数学模型定量解析等方法，从药理作用、体内过程、多组分和药效的定量关系三方面，探讨多组分的协同作用，可为EGB原料药及制剂质量控制提供依据，也可为中药多组分研究提供思路。

多组分协同作用是中药发挥整体药效的关键，但其协同作用机制有待探究。前期研究发现，多组分相互作用可影响体内动力学过程，推测体内过程改变是其协同作用机制之一。如何定量解析多组分体内行为和药效间的关系，是中药研究的难点之一。本项目主要研究内容：银杏黄酮和银杏内酯相互作用对脑缺血相关药理指标的影响；多组分相互作用对多组分体内过程的影响；结构方程模型对银杏叶提取物抗脑缺血的协同作用进行解析。结果表明，银杏黄酮和银杏内酯在体外抗氧化实验中表现出协同作用，以银杏黄酮:银杏内酯4:1时最强；银杏黄酮和银杏内酯可通过清除自由基、抑制炎症反应等改善大鼠脑缺血再灌注损伤，两者合用可协同起效；银杏黄酮和银杏内酯联合用药可使银杏内酯A和银杏内酯B在体内的吸收速度加快；结构方程模型解析结果表明，银杏叶提取物各组分对SOD、MDA、IL及大脑氨基酸水平表现出协同作用，以芦丁和银杏内酯A、银杏内酯C和异鼠李素、银杏内酯A、B、C之间、以及芦丁和芹菜素之间协同作用最为明显。