DataSheet_1_Systems Pharmacology Approach to Investigate the Mechanism of Kai-Xin-San in Alzheimer’s Disease.xlsx

Alzheimer's disease (AD) is a complex neurodegenerative disease characterized by cognitive dysfunction. Kai-Xin-San (KXS) is a traditional Chinese medicine (TCM) formula that has been used to treat AD patients for over a thousand years in China. However, the therapeutic mechanisms of KXS for treating AD have not been fully explored. Herein, we used a comprehensive network pharmacology approach to investigate the mechanism of action of KXS in the treatment of AD. This approach consists of construction of multiple networks and Gene Ontology enrichment and pathway analyses. Furthermore, animal experiments were performed to validate the predicted molecular mechanisms obtained from the systems pharmacology-based analysis. As a result, 50 chemicals in KXS and 39 AD-associated proteins were identified as major active compounds and targets, respectively. The therapeutic mechanisms of KXS in treating AD were primarily related to the regulation of four pathology modules, including amyloid beta metabolism, tau protein hyperphosphorylation process, cholinergic dysfunction, and inflammation. In scopolamine-induced cognitive dysfunction mice, we validated the anti-inflammatory effects of KXS on AD by determining the levels of inflammation cytokines including interleukin (IL)-6, IL-1β, and tumor necrosis factor (TNF)-α. We also found cholinergic system dysfunction amelioration of KXS is correlated with upregulation of the cholinergic receptor CHRNB2. In conclusion, our work proposes a comprehensive systems pharmacology approach to explore the underlying therapeutic mechanism of KXS for the treatment of AD.

阿尔茨海默病（Alzheimer's disease, AD）是一种以认知功能障碍为特征的复杂神经退行性疾病。开心散（Kai-Xin-San, KXS）是一种中药方剂（traditional Chinese medicine, TCM），在中国已有千余年用于治疗AD患者的历史，但其治疗AD的具体机制尚未完全阐明。本研究采用整合网络药理学方法，探究KXS治疗AD的作用机制。该方法涵盖多网络构建、基因本体（Gene Ontology, GO）富集分析及通路分析。此外，本研究通过动物实验验证了基于系统药理学分析得到的预测分子机制。研究共鉴定出KXS中的50种化学成分与39个AD相关蛋白，分别作为主要活性成分与作用靶点。KXS治疗AD的潜在机制主要与四大病理模块的调控相关，包括β淀粉样蛋白代谢、tau蛋白过度磷酸化过程、胆碱能系统功能障碍以及炎症反应。在东莨菪碱诱导的认知功能障碍小鼠模型中，本研究通过检测白细胞介素（interleukin, IL）-6、IL-1β及肿瘤坏死因子（tumor necrosis factor, TNF）-α等炎症细胞因子水平，验证了KXS的抗AD抗炎作用。同时发现，KXS对胆碱能系统功能障碍的改善作用与胆碱能受体CHRNB2的上调存在相关性。综上，本研究提出了一套完整的系统药理学方法，用以探究KXS治疗AD的潜在分子机制。