基于微透析在线分析技术和代谢组学方法的人参远志配伍治疗阿尔茨海默病的药效物质基础和作用机制研究

Alzheimer’s disease (AD) is an age-related disease characterized in cognitive declining. The traditional Chinese medicine has significant application for treatment of AD. However, it is quite difficult to study their overall mechanism of action because of its complex composition and multi targets. It is a hot spot to develop natural medicine to treat AD. In this study, traditional Chinese medicine Ginseng and Polygala tenuifolia were combined as compatibilities. We employed an online method to combine microdialysis technology and liquid chromatography -mass spectrometry, established a pharmacokinetics-pharmacodynamics model to research pharmacokinetics rules of compatibilities metabolites group in the blood and brain of AD rats, in order to explain the pharmacodynamic material basis of compatibilities for treating AD. We applied a metabolomics method for determining the potential biomarkers to study the overall mechanism of compatibilities for treating AD and the reasonable compatibility proportion. The online combined microdialysis -liquid chromatography -mass spectrometry method with the advantage of samping in situ, monitoring detection and no sampling pretreatment is expected to provide strong technical support for studing the overall mechanism of action for treating AD of the traditional Chinese medicine.

阿尔茨海默病（Alzheimer’s disease，AD）是一种以认知功能减退为主要症状的老年性疾病，中药在治疗AD上历史悠久，然而由于中药成分复杂、作用靶点多样，限制了其在治疗AD中的应用，开发中药治疗AD是当今热点之一。本项目将传统治疗健忘痴呆的中药人参和远志配伍，建立微透析-超高效液相色谱/质谱在线联用分析方法，借助药动学-药效学模型，研究人参远志配伍药对入血、入脑代谢物组在AD大鼠体内的药代动力学规律，旨在解释人参远志配伍治疗AD的药效物质基础；并借助代谢组学研究方法考察相关内源性生物标志物的变化，阐明人参配伍治疗AD的合理比例和整体作用机制。本研究充分发挥微透析技术原位取样、无需样品预处理、动态检测等优点，建立的微透析超高效液相色谱/质谱在线联用分析方法有望对中药治疗AD的研究提供强有力的技术支撑。

阿尔茨海默病（Alzheimer’s disease，AD）是一种以认知功能减退为主要症状的老年性疾病，中药在治疗AD上历史悠久，然而由于中药成分复杂、作用靶点多样，限制了其在治疗AD中的应用。本项目将传统治疗健忘痴呆的中药人参和远志配伍，应用所建立的在线微透析-LC-MS/MS联用分析方法成功分析了人参、远志对AD大鼠神经递质的影响，结果表明经过人参治疗后大鼠脑海马谷γ-氨基丁酸（GABA）、甘氨酸（Gly）和乙酰胆碱（Ach）含量增加，谷氨酸（Glu）、天冬氨酸（Asp）含量减少，皮层和血浆Ach和5-羟色胺（5-HT）含量增加。经远志治疗后大鼠脑海马Glu和GABA含量降低，Glu/GABA比值降低，GABA、Gly、Ach、5-HT和DA含量升高。分析了人参、远志的化学成分，其中鉴定人参化合物44个，远志化合物136个，并在此基础上解释了人参远志配伍治疗AD的药效物质基础，共鉴定了113个人参化合物及代谢产物入血成分，其中在大鼠的血浆、肝脏和大脑组织中鉴定了53、55和6个，31个是通过氧化作用得到；共鉴定了48个远志化合物及代谢产物入血成分，其中在大鼠的血浆、肝脏和大脑组织中粪便鉴定了13、32和3个，同时在大鼠尿样和粪便中鉴定了40和73个。研究了参远志配伍药在AD大鼠体内的药代动力学规律，与人参单药相比，人参与远志配伍给药后，Tmax减小，T1/2，Cmax 和AUC增大。与远志单药相比，人参远志配伍给药后， Tmax, T1/2 增加, Cmax减小，表明人参远志配伍前后各类化合物在大鼠体内的药代动力学发生了显著的改变。借助代谢组学研究方法得出人参远志配伍治疗AD的机制与影响34种生物标志物和乙醛和二羧酸代谢，果糖和甘露糖代谢，氨基糖和苷糖代谢，柠檬酸代谢，色氨酸代谢，精氨酸和脯氨酸代谢，甘油磷脂代谢，丙氨酸、天冬氨酸和谷氨酸代谢，卟啉和叶绿素代谢，半乳糖代谢，牛磺酸和亚牛磺酸代谢和维生素6代谢等通路有关。