基于Aβ产生和突触功能调节研究开心散加味新方治疗阿尔兹海默病的作用机制

Kai-Xin-San, a famous classical prescription in traditional Chinsese medicine (TCM), has been proved with clear effects in treating Alzheimer's disease (AD). Among the pathological characteristics of AD, the blood stasis is obviously one aspect of the pathological features of this disease, but Kai-Xin-San is lack of efficacy in promoting blood circulation and removing blood stasis. Previously, a new prescription, the Modified Kai-Xin-San, was set up by us by adding several traditional Chinese herbs to Kai-Xin-San, these added Chinese herbs were identified with the efficacy in removing blood stasis and with the activity in treating AD. Our previous results showed that, compared with Kai-Xin-San, the novel Modified Kai-Xin-San showed more efficacy in treating AD, the content levels of the active ingredients with the anti-AD activity in Kai-Xin-San were also found increased in the brains of the Modified Kai-Xin-San-treated animals, which indicates that, the modified new prescription could improve the bioavailability of the original prescription. In this project, according to the amyloid cascade hypothesis in AD pathology, an AD mouse model which carrying multiple mutation genes in AD patients and two kinds of in vitro cultured cells with the pathological characteristics of the AD were used to study the effects of Kai-Xin-San and the Modified Kai-Xin-San in treating AD. Besides these, their underlying mechanisms based on the modulation of the Aβ production and the regulation of the synaptic function will also be investigated in this project. In addition, the in vivo molecular and functional imaging technologies will also be applied in this project in order to further clarify the characteristics of the effects of these prescriptions in treating AD. These above researches may provide us useful references and the experimental basis for AD treatment in TCM clinic and for further research and development of new TCM drugs.

经典名方开心散具有明确的治疗老年痴呆症（阿尔兹海默病，AD）作用，临床和实验研究表明，AD存在明显的血瘀病理特征，但开心散原方活血祛瘀之力偏弱。我们前期选择具有活血祛瘀功效且具有抗AD活性的中药加入开心散组成开心散加味新方，研究表明开心散加味后抗AD作用增强，且能促进开心散原方抗AD活性成分在脑内的分布，提高原方的生物利用度。本项目拟采用携带多基因突变位点的AD小鼠模型，并采用体外培养的AD病理细胞模型，进一步对比研究上述两方的抗AD作用，并基于Aβ级联假说，进一步选择Aβ产生的始动环节与最直接反映AD认知功能改变的突触结构和功能调节的较终末环节两个关键节点，同时结合活体分子及功能影像学技术，较系统地研究上述两方治疗AD的不同作用特点和机理，从而从上述角度阐明开心散活血化瘀加味治疗AD的配伍内涵，为临床AD的中医药治疗和进一步中药新药研发提供现代生物医学依据和前期实验研究基础。

经典名方开心散作为一首治疗老年痴呆症（阿尔兹海默病，AD）的基础方在临床广为使用， AD临床上存在明显的“虚”“痰”“瘀”病理特征，开心散原方补气健脾、豁痰开窍之效强，但补肾填精、活血祛瘀之力弱。我们在开心散原方基础上加味具有活血祛瘀等功效的中药组成开心散加味新方，进一步采用携带多基因突变位点的AD小鼠模型和AD病理细胞模型，对比研究了开心散原方及加味新方对AD的治疗作用，并基于Aβ产生和突触功能调节探讨了相关机制。我们首先制备了开心散原方及开心散加味新方药物制剂，进行了化学成分和制剂稳定性研究，并对关键成分进行了网络药理学分析，结果表明，开心散原方及加味新方制剂具有良好的稳定性，检测的24种化合物可作为质量控制的成分。行为学研究表明，开心散及加味新方治疗均能不同程度改善5×FAD小鼠的学习记忆能力；电生理学研究表明，开心散及加味新方均能不同程度改善5×FAD小鼠海马脑片CA3-CA1通路长时程增强现象，并对兴奋性及抑制性突触传递具体不同程度的改善作用；形态学等研究表明，开心散及加味新方均可以显著改善5×FAD小鼠海马神经元突触结构和脑部微小血管病变等；进一步的小动物活体fMRI及[18F]FDG PET-CT影像学研究表明，开心散原方及加味新方均能不同程度改善5×FAD小鼠海马脑能量代谢等。在体及细胞学实验还表明开心散原方及加味新方均能影响AD小鼠及细胞模型的Aβ产生和沉积。开心散加味新方的上述作用均明显优于开心散原方，表明活血化瘀等加味有助于改善经典名方开心散对AD的治疗作用，并有助于改善对AD模型Aβ产生、突触结构和功能的调节作用。上述研究从阿尔兹海默病Aβ产生及突触结构和功能调节的角度，阐明了经典名方开心散及其活血化瘀等加味治疗阿尔兹海默病的现代生物医学内涵，为临床阿尔兹海默病的中医药治疗，为进一步中药新药研发提供了相关实验研究基础。