基于中医方证代谢组学的生脉散干预老年痴呆症大鼠的药效物质基础研究

A Research for Evaluating the Effectiveness and Discovering the Pharmacodynamic Substances of Sheng Mai San(SMS) in the Treatment of Alzheimer's Disease(AD)

目的:本实验主要利用中医方证代谢组学技术评价生脉散预防老年痴呆的药效及发现其关键物质基础。方法:首先采用灌胃三氯化铝联合腹腔注射D-半乳糖连续90天建立老年痴呆大鼠模型,并利用UPLC-HDMS技术采集和分析模型大鼠尿液代谢数据,共确定了42个老年痴呆症的生物标记物;同时干预组在造模同时采用连续给与生脉散冻干粉溶液(5.3 g·kg-1)对模型大鼠进行治疗。结果:通过Morris水迷宫、脑组织病理学检测及Aβ免疫组化等方法评价了生脉散的干预作用,发现了生脉散具有抑制Aβ毒性、保护神经元和改善认知障碍的作用;在此基础上利用代谢组学技术评价了干预组对42个老年痴呆生物标志物的调节作用,发现生脉散主要影响了色氨酸代谢和脂质代谢的多个靶点;同时采用血清药物化学方法研究了生脉散的体内直接作用物质,找到30种成分并与生脉散明显回调的生物标志物相关联。结论:本实验发现了五味子木质素类及人参皂苷类的多种成分与色氨酸代谢和脂质代谢极度相关,为生脉散预防老年痴呆的药效物质基础。

The aim of this study was to evaluate the effectiveness and discover the pharmacodynamic substances of SMS in the treatment of AD. AD model rats were successfully established with the 90-day continuous intragastric administration of aluminium chloride and intraperitoneal injection of D-galactose. Morris water maze test, histopathological staining and immunohistochemical staining were adopted to evaluate the progress of AD in model rats and the efficacy of SMS. Metabolomic technology platform was used to reveal the potential biomarkers and the metabolic network of AD and the target spots of impacted metabolic pathways after the administration of SMS. As a result, it was found that SMS performed the regulatory effects on certain metabolic pathways, in which tryptophan and lipid metabolism were involved in the main metabolic pathways influenced by SMS. Serum pharmacochemical technologies of TCM were introduced to investigate the constituents absorbed into the blood after lavage administration of SMS. In conclusion, the constituents were associated with target spots of influenced metabolic pathways which elucidated the pharmacodynamic basis. The components of lignins and their metabolites and ginsenoside were the most relevant materials of AD associated with tryptophan and lipid metabolism, which may be the pharmacodynamic basis of AD playing therapeutic roles of SMS.