开心散与生脉散抗阿尔茨海默症“同病异治”作用机制网络药理学分析与中枢神经免疫调控效用验证

Network Pharmacological Analysis of the Mechanism of Kai-Xin-San and Sheng-Mai-San Against Alzheimer’s Disease with "Same Disease with Different Treatments" and Effect Verification of Central Nervous Inflammation Regulation

目的基于网络药理学方法研究开心散(KXS)与生脉散(SMS)“同病异治”抗阿尔茨海默症作用机制,并通过整体动物与离体细胞实验进行验证。方法通过数据库搜集筛选开心散与生脉散复方所含药味化学成分并预测其潜在靶点,构建复方-中药-成分-靶点网络。筛选复方共有与优势靶点,构建蛋白互作网络并进行拓扑分析,进行KEGG通路和GO功能富集分析。基于网络药理学分析获得的开心散与生脉散共性与个性的生物学GO分析结果,分别建立小鼠海马Aβ注射拟阿尔茨海默症痴呆小鼠模型与小鼠小胶质BV2细胞炎性因子损伤模型,检测相应指标,采用行为学结合生化指标分析验证网络药理学分析结果。结果筛选出开心散与生脉散复方共有成分有109个,共有靶点490个,抗AD潜在共性调控靶点375个。开心散抗AD潜在优势调控靶点92个,生脉散抗AD潜在优势调控靶点90个。淀粉样蛋白代谢与转运、cAMP与AMPK信号通路、突触功能调控等为开心散与生脉散抗AD潜在共性调控信号通路与生物学事件。NF-κB等为开心散抗AD优势调控信号通路与生物学事件;趋化因子、胆碱能突触等为生脉散抗AD优势调控信号通路与生物学事件。验证实验结果显示开心散与生脉散能够有效改善Aβ海马区注射小鼠认知功能障碍,降低小鼠海马和小胶质细胞的炎性因子表达。结论开心散与生脉散能通过共有及各自优势靶点发挥抗AD作用。

Objective To study the anti-Alzheimer’s disease(anti-AD) mechanism of "treating the same disease differently" of Kai-Xin-San(KXS) and Sheng-Mai-San(SMS) based on the network pharmacology method, and to verify the whole animal and in vitro cell experiments.Methods The chemical components of KXS and SMS were collected and screened through database, and their potential targets were predicted to build the network of compound-traditional Chinese medicine-component-target. The common and dominant targets of the compound were screened. The protein interaction network was constructed and the topological analysis was carried out, and the KEGG pathway and GO function enrichment were analyzed. Based on the biological GO analysis results of commonness and individuality of KXS and SMS obtained by network pharmacology analysis, the mice hippocampus Aβ injection to AD dementia model in mice and mice BV2 microglia inflammatory factor damage model were established respectively. The corresponding indexes were detected, and the results of network pharmacology analysis were verified by behavioral analysis combined with biochemical index analysis.Results There were 109 common components, 490 targets and 375 anti-AD targets of KXS and SMS. There were 92 dominant anti-AD targets in KXS and 90 dominant anti-AD targets in SMS. Amyloid metabolism and transport, cAMP and AMPK signaling pathways, and synaptic function regulation were the potential common regulatory signaling pathways and biological events of KXS and SMS against AD. NF-κB and other factors were the dominant signal pathway and biological events of anti-AD in KXS. Chemokines, cholinergic synapses, etc., regulated the signal pathways and biological events of the anti-AD dominance of SMS. The verification results showed that both them could effectively improve the cognitive dysfunction of Aβ-injected mice and reduce the expression of inflammatory factors in the hippocampus and BV2 microglia cell of mice. Conclusion KXS and SMS can exert anti-AD effects through common and respective dominant targets.