基于网络药理学和实验验证远志-石菖蒲药对治疗阿尔茨海默病的作用机制

Exploration on mechanism of Polygalae Radix and Acori Tatarinowii Rhizoma in treating Alzheimer′s disease based on network pharmacology and experimental verification

基于网络药理学、分子对接及动物实验探讨远志-石菖蒲药对防治阿尔茨海默病(Alzheimer′s disease,AD)的作用靶点及其作用机制。该研究通过GeneCards等数据库收集AD疾病相关靶点,从TCMSP等数据库获得远志-石菖蒲的主要活性成分及其对应的靶点,运用Cytoscape软件绘制“药物-活性成分-作用靶点-疾病”网络及采用STRING数据库建立其共有靶点蛋白互作(PPI)网络。随后分别利用DAVID及Metascape数据库进行GO功能富集分析和KEGG通路富集分析。通过AutoDock Vina软件对远志-石菖蒲的主要活性成分与潜在作用核心靶点进行分子对接。采用体内实验验证远志-石菖蒲对东莨菪碱(scopolamine,SCOP)模型小鼠认知功能的影响。结果筛选出远志-石菖蒲具有以山柰酚(kaempferol)、氧杂蒽酮(onji-xanthone I)、异紫花前胡内酯(marmesin)为主的7个活性成分,涉及56个潜在治疗靶点,其中以AKT1、PTGS2、TNF等为核心靶点,与NF-κB炎症通路密切相关。分子对接结果表明网络药理学所预测核心靶点能与远志-石菖蒲主要活性成分稳定结合。新物体识别实验结果提示远志-石菖蒲可改善SCOP诱导的小鼠学习记忆能力。病理切片及相关试剂盒检测提示远志-石菖蒲能减轻其中枢胆碱能神经元损伤,同时改善SCOP模型小鼠的抗氧化能力。Western blot表明远志-石菖蒲可下调SCOP模型TLR4、NF-κB及相关炎症因子(TNF-α、IL-1β、IL-6)的蛋白表达水平。该研究表明远志-石菖蒲在阿尔茨海默病的治疗方面凸显出多成分、多靶点、多途径协同作用的特点,为临床应用提供了科学性的实验依据。

This study aimed to explore the action targets and mechanisms of Polygala tenuifolia and Acorus tatarinowii in treating Alzheimer′s disease(AD)based on network pharmacology,molecular docking,and animal tests.The AD-related targets were collec-ted from GeneCard and the main active ingredients and targets of P.tenuifolia and A.tatarinowii from the TCMSP.Cytoscape was applied to construct the"Chinese herb-active ingredient-target-disease"network,followed by the construction of protein-protein interaction(PPI)network using STRING.GO biological function and KEGG pathway enrichment analysis was performed by DAVID and Metascape.The main active components of P.tenuifolia and A.tatarinowii and their potential core targets were docking using AutoDock Vina.The effects of P.tenuifolia and A.tatarinowii on the cognitive function were verified in mice with scopolamine(SCOP)-induced cognitive impairment.A total of seven active ingredients including kaempferol,onjixanthoneⅠ,and marmesin and 56 potential targets of P.tenuifolia and A.tatarinowii were screened out,with the core targets covering AKT1,PTGS2,TNF,and NF-κB inflammation pathway mainly involved.The results of molecular docking also showed that the main active components of P.tenuifolia and A.tatarinowii stably bond to the core targets predicted by network pharmacology.The new object recognition experiment suggested that P.tenuifolia and A.tatarinowii improved the learning and memory abilities of mice after SCOP induction.As revealed by pathological section observation and relevant kit assay,P.tenuifolia and A.tatarinowii reduced the damage of central cholinergic neurons and enhanced the antioxidant ability of SCOP-induced mice.Western blot confirmed that P.tenuifolia and A.tatarinowii down-regulated the protein expression levels of TLR4,NF-κB,and related inflammatory factors(TNF-α,IL-1β,and IL-6).All these have suggested that P.tenuifolia and A.tatarinowii inhibits AD via multiple components,multiple targets,and multiple pathways,which has provided an experimental basis for the clinical application of P.tenuifolia and A.tatarinowii for the treatment of AD.