基于BAP理论从细胞内Aβ清除和生物利用度角度探讨当归芍药散对阿尔茨海默病的作用机制研究

Dangdui-Shaoyao-San (DSS) was found to improve cognitive dysfunction of patients and model animals of Alzheimer's disease (AD).The pathogenesis of Chinese medicine for the dementia was considered as "deficiency in origin and excess in superficiality"."Deficiency in origin" means kidney essence deficiency and insufficiency of Qi-Blood, while "Excess in superficiality" means phlegm retention in head and Qi Blood stasis. DSS is capable of "reinforce insufficiency and reduce excessiveness". Our preceding research found that DSS could decrease the content of β-amyloid protein (Aβ) in the brain of senescence accelerated mouse, and enhance the bioavailability of Levistilide, a effective component of DSS, afer oral administration of DSS. Basied on the theory that being absorbed to reach a certain concentration is the first step of drug action and in the guidelines of bioethnopharmaceutical analytical pharmacology (BAP) research strategy, our team will analyze the compound of DSS in blood to discover the absorbed bioactive compound (ABC) of DSS; study the effect of DSS on the bioavailability of it's ABC; discover the effect of DSS and its ABC on the efflux of intracellular Aβ to protect mitochondria dysfunction against Aβ's toxicity (there is still no paper about the effect of DSS on Aβ which is the leading cause of AD). Our study will reveal the mechanism of DSS on AD basied on the effects of drug on the body as well as that of body on the drug, which is innovative on the ideas and pharmacological mechanism and will provide new experimental basis to prove the scientificity and rationality of Traditional Chinese Medicine.

研究发现当归芍药散（DSS）能改善阿尔茨海默病（AD）患者和动物的认知功能障碍，痴呆病机为本虚标实，本虚为肾精亏损和气血不足，标实为痰浊阻窍和气滞血瘀，DSS具有补本虚、祛标实特点。本课题前期研究发现DSS能减少动物脑内β淀粉样蛋白（Aβ）含量，并能提高其中的有效成分欧当归内酯的生物利用度。药物被机体吸收达到一定浓度是其发挥药效的前提，本课题运用生物方剂分析药理（BAP）研究策略，分析DSS的血中转移成分，寻找DSS中吸收的生物活性成分（ABC）,探讨DSS对其ABC生物利用度的影响，并从促进细胞内Aβ清除及调节Aβ导致线粒体功能紊乱角度，深入探讨DSS及其ABC的神经保护作用机制（Aβ为AD发病核心因素，尚无DSS对其作用报道）。本课题既从药物对机体，又从机体对药物作用角度，较全面地诠释DSS抗AD药理作用机制，在研究思路与内容上均有创新性，为中药复方组方科学性和合理性提供新的实验依据。

本课题既从药物对机体，又从机体对药物作用角度，较全面地诠释当归芍药散抗阿尔茨海默病（AD)的药理作用机制。结果显示芍药内酯苷、芍药苷、苯甲酸、没食子酸、阿魏酸、欧当归内酯A、川芎内酯I、川芎内酯A、16-羰基泽泻醇A、3β-乙酰氧基苍术酮、芍药苷元酮、泽泻醇 B、丁烯基苯酞、环氧泽泻烯、芍新苷、丹皮酚、白术内酯Ⅲ、茯苓酸、儿茶素、白术内酯Ⅱ很可能为当归芍药散的入血成分。当归芍药散主要入血成分藁本内酯、芍药内酯苷、阿魏酸组对AD动物模型APP/PS1小鼠在学习记忆能力上具有改善作用。当归芍药散入血成分藁本内酯、芍药内酯苷对AD动物模型APP/PS1小鼠脑内海马CA1区线粒体损伤有一定的改善作用，保护Aβ造成的线粒体损伤，其机制可能是通过改善Aβ造成的线粒体损伤从而起到神经保护作用。体外研究结果表明，25 μM藁本内酯和100 μM芍药内酯苷对线粒体的保护效果较为明显。当归芍药散的主要入血成分欧当归内酯A、丁烯基苯酞、泽泻醇B的Cmax和AUC显著高于单体丁烯基苯酞、泽泻醇B。当归芍药散对方中主要入血成分吸收的机制研究表明，阿魏酸（FA）在复方中有着更好的生物利用度，其机制可能是由于复方中的其它成分可以同时抑制外排蛋白和抑制CYP3A4的活性，对增加FA的生物利用度起到一种协同作用。本项目在研究思路与方法上，为中药复方研究提供了一条新的思路。