基于网络药理学与分子对接探讨风引汤治疗癫痫作用机制  

Mechanism of Fengyin Tang in Treating Epilepsy Based on Network Pharmacology and Molecular Docking

作　　者：邱璐琦 丁舒飞 蒋东晓 阮叶萍[3] QIU Luqi;DING Shufei;JIANG Dongxiao;RUAN Yeping

机构地区：[1]浙江中医药大学附属第三医院中药药剂科,浙江杭州310009 [2]绍兴市中医院中药药剂科,浙江绍兴312000 [3]浙江中医药大学药学院,浙江杭州310053

出　　处：《新中医》2022年第15期6-14,共9页New Chinese Medicine

基　　金：浙江省自然科学基金项目(LY20H280004)。

摘　　要：目的:基于网络药理学与分子对接探讨风引汤治疗癫痫的潜在作用机制,并通过动物实验验证其预测靶点的可靠性。方法:结合文献,借助TCMSP、CTD数据库,获取风引汤的活性成分及其相关分子靶点;通过GeneCards、DisGeNET、TTD和OMIM数据库检索癫痫相关的基因,运用Cytoscape3.7.0软件构建风引汤治疗癫痫的蛋白相互作用网络和成分-靶点网络,筛选出风引汤治疗癫痫的关键靶点;利用R软件将关键靶点进行GO功能和KEGG通路富集分析;通过分子对接进行半柔性对接反向验证并运用实时荧光定量PCR法对关键靶点进行验证。结果:筛选出风引汤治疗癫痫的活性成分134个,与癫痫的共有靶点46个,其中风引汤抗癫痫的关键靶点主要包括半胱氨酸天冬氨酸蛋白酶3(CASP3)、转录因子AP-1(JUN)、Myc原癌基因蛋白(MYC)、前列腺素G/H合成酶2(PTGS2)和过氧化氢酶(CAT)等,涉及信号通路有AGE-RAGE信号通路、p53信号通路、神经退行性信号通路等。实时荧光定量PCR实验证明风引汤能降低癫痫大鼠海马中CASP3、JUN mRNA表达(P<0.05),升高CAT mRNA表达(P<0.05)。结论:风引汤治疗癫痫主要作用靶点与CASP3、JUN、CAT有关,其作用机制可能与AGE-RAGE、p53、神经退行性等信号通路相关。Objective:To explore the potential mechanism of Fengyin tang in treating epilepsy based on network pharmacology and molecular docking,and to verify the reliability of its prediction target through animal experiments.Methods:The active ingredients and related molecular targets of Fengyin tang were obtained with the help of literatures and TCMSP and CTD databases.GeneCards,DisGeNET,TTD and OMIM databases were used to search the epilepsy related genes,and Cytoscape3.7.0 software was used to construct the protein interaction network and component-target network of Fengyin tang in treating epilepsy,and the key targets of Fengyin tang in treating epilepsy were screened out.Key targets were carried out enrichment analysis for GO function and KEGG pathway via R software.Reverse verification of semi-flexible docking was performed by molecular docking,and the key targets were verified by Quantitative Real-time PCR.Results:A total of 134 active components of Fengyin tang in treating epilepsy were screened out,and 46 common targets with epilepsy were identified.The key antiepileptic targets of Fengyin tang mainly included Caspase-3(CASP3),activator protein AP-1(JUN),Myc proto-oncogene protein(Myc),prostaglandin G/H synthase-2(PTGS2)and catalase(CAT).The involved signaling pathways included AGE-RAGE signaling pathway,p53 signaling pathway,neurodegenerative signaling pathway,etc.Quantitative Real-time PCR showed that Fengyin tang could decrease the mRNA expression of CASP3 and JUN and increase the mRNA expression of CAT in hippocampus of epileptic rats(P<0.05).Conclusion:The main targets of Fengyin tang in treating epilepsy are related to CASP3,JUN and CAT,and its mechanism may be related to AGE-RAGE,p53 and neurodegeneration signal pathways.

关 键 词：癫痫 风引汤 网络药理学 分子对接 

分 类 号：R742.1[医药卫生—神经病学与精神病学]