摘要
目的 基于网络药理学分析半夏白术天麻汤治疗癫痫的作用靶点、信号通路和潜在作用机制,并通过分子对接技术和动物实验进行验证。方法 通过BATMAN-TCM等数据库筛选半夏白术天麻汤的活性成分及药物靶点,通过GeneCards等数据库获取癫痫相关疾病靶点,取交集靶点;构建“中药-活性成分-靶点-疾病”网络;构建PPI网络并筛选核心靶点;进行GO、KEGG功能富集分析;构建“通路-靶点-活性成分”网络并筛选核心活性成分。运用分子对接技术对核心靶点与核心活性成分进行验证;在动物实验中,以氯化锂-匹罗卡品诱导癫痫大鼠模型,并将40只Wistar大鼠分为正常对照组、模型对照组、卡马西平组、半夏白术天麻汤组,通过行为学观察癫痫发作情况,尼氏染色观察海马组织神经元损伤,免疫组化检测BAX、BCL-2、Caspase-3蛋白表达。结果 筛选出1072个半夏白术天麻汤的药物靶点,筛选出1046个癫痫的疾病靶点,半夏白术天麻汤治疗癫痫的220个交集靶点。获取PPI网络核心靶点AKT1、ALB、ACTB、INS、TNF;KEGG通路主要涉及TNF通路、IL-17信号通路、cAMP信号通路、神经退行性变的途径-多重疾病、含血清素的神经突触、多巴胺能神经突触等通路;“通路-靶点-活性成分”网络中相关性最高的核心活性成分为白桦脂醇(Betulin)、麻黄碱(Ephedrine)、麦角胺(Ergotamine)、百里香酚(Thymol);分子对接结果显示核心靶点与核心活性亲和力良好。动物实验发现半夏白术天麻汤能有效降低大鼠痫性发作,改善大鼠海马神经元损伤,显著降低神经元凋亡百分率,显著下调促凋亡蛋白BAX、Caspase-3表达,上调抗凋亡蛋白BCL-2表达趋势。结论 半夏白术天麻汤治疗癫痫具有多成分、多靶点、多途径的特点,抑制神经元凋亡减少海马神经元损伤可能是其治疗癫痫的重要机制之一。
Objective To analyze the target,signal pathway and potential mechanism of Banxia Baizhu Tianma Decoction in the treatment of epilepsy based on network pharmacology,and to verify it by molecular docking technology and animal experiments.Methods The active ingredients and drug targets of Banxia Baizhu Tianma Decoction were screened by BATMAN and other databases.The targets of epilepsy-related diseases were obtained by GeneCards and other databases,and the intersection targets were taken.Constructing‘drug-ingredient-target-disease’network and PPI network to screen the core targets.The core active ingredients were screened according to GO,KEGG functional enrichment analysis and′pathway-target-active ingredient′network.Molecular docking was used to verify the core targets and core active ingredients.In the animal experiment,the rat model of epilepsy was induced by lithium chloridepilocarpine,and 40 Wistar rats were divided into normal control group,model control group,carbamazepine group and Banxia Baizhu Tianma Decoction group.The seizures were observed by behavior.Nissl staining was used to observe neuronal damage in hippocampus.Immunohistochemistry was used to detect the expression of BAX,BCL-2 and Caspase-3 protein.Results A total of 1072 targets of Banxia Baizhu Tianma Decoction were screened,1046 disease targets of epilepsy were screened,and 220 intersection targets of Banxia Baizhu Tianma Decoction in the treatment of epilepsy were screened.The core targets AKT1,ALB,ACTB,INS and TNF of PPI network were obtained.KEGG pathway mainly involves TNF signaling pathway,IL-17 signaling pathway,cAMP signaling pathway,pathways of neurodegeneration-multiple diseases,serotonergic synapse and Dopaminergic synapse.The core active ingredients with the highest correlation in the′pathway-target-active component′network were Betulin,Ephedrine,Ergotamine and Thymol.Results of molecular docking indicated that the core target had satisfactory affinity with those active ingredients.Animal experiments showed that Banxia Baizhu Tianma Decoction could effectively reduce epileptic seizures in rats,improve hippocampal neuronal damage in rats,significantly reduce the percentage of neuronal apoptosis,significantly down-regulate the expression of pro-apoptotic proteins BAX and Caspase-3,and up-regulate the expression of antiapoptotic protein BCL-2.Conclusion Banxia Baizhu Tianma Decoction has the characteristics of multi-component,multi-target and multi-pathway in the treatment of epilepsy.Inhibiting neuronal apoptosis and reducing hippocampal neuronal damage may be one of the important mechanisms for its treatment of epilepsy.
作者
杨欣
符晋
蒋萃
开胤桦
何佳忆
郭向鑫
田茸
YANG Xin;FU Jin;JIANG Cui;KAI Yinhua;HE Jiayi;GUO Xiangxin;TIAN Rong(Chengdu University of Traditional Chinese Medicine,Chengdu 611137,China)
出处
《世界科学技术-中医药现代化》
北大核心
2025年第3期776-791,共16页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
基金
国家自然科学基金委员会面上项目(81973929):熄风化痰法抑制癫痫风痰证候神经元凋亡及circRNA-miRNA-mRNA调控网络机制研究,负责人:田茸
四川省自然科学基金委员会青年基金项目(2023NSFSC1805):基于miR142/PINK1/Parkin通路促进癫痫大鼠海马线粒体自噬探讨熄风化痰法抑制癫痫反复发作机制,负责人:蒋萃
成都中医药大学杏林人才苗圃项目(MPRC2022003):基于miR142/PINK1通路促进癫痫大鼠海马线粒体自噬探讨熄风化痰法抑制癫痫反复发作机制,负责人:蒋萃。
关键词
半夏白术天麻汤
癫痫
网络药理学
分子对接
Banxia Baizhu Tianma Decoction
Epilepsy
Network pharmacology
Molecular docking
