摘要
为了解诃子治疗的潜在靶点及相关作用机制,试验采用网络药理学、分子对接等方法,联合牦牛源沙门菌攻毒试验验证.通过HERB组鉴数据库、TCMSP数据库和SwissTargetPrediction网页工具筛选诃子中的活性成分以及潜在作用靶点,通过OMIM和GeneCards数据库以"胃肠炎"为关键词进行搜索,使用Cytoscape和STRING数据库构建诃子PPI网络筛选出关键靶点,通过Venny平台获得诃子与肠炎之间的交集靶点,通过DAVID数据库进行基因本体论(GO)、京都基因和基因组百科全书数据库(KEGG)途径富集分析,对筛选的核心靶点进行分子对接验证;之后建立小鼠胃肠炎症模型,观察胃肠道组织病理变化,并利用免疫印迹试验(Western blot)验证诃子对靶点蛋白的影响.结果显示:分析整理了诃子8个主要活性成分,筛选到了诃子靶点118个,获取胃肠炎靶点11 161个,交集靶点100个;通过PPI网络得到了诃子与肠炎疾病的PTGS2、CASP3、SLC3A2、Bax、Bcl-2、TP53核心靶点;GO与KEGG富集分析分别收获337和138条目,主要关联趋化因子通路、PI3K-Akt信号通路、细胞凋亡相关通路、铁离子转运相关通路、NF-κB信号通路等;分子对接结果显示,诃子排名第一活性成分诃子次酸可以与Bax、Bcl-2、PTGS2、SLC3A2靶点很好地结合,主要通过氢键、疏水作用、π-π堆积力等分子间作用力;病理组织切片显示,诃子可以显著恢复胃肠道组织损伤;Western blot试验结果显示,诃子可以通过Bax/Bcl-2和PTGS2/SLC3A2通路来调节细胞凋亡和铁死亡过程,达到治疗肠道炎症损伤的效果.结果表明,诃子可以通过Bax/Bcl-2和PTGS2/SLC3A2通路调节细胞凋亡和铁死亡过程来调控肠炎的发生发展,诃子治疗肠炎疾病具有多靶点和多通路的特点.
Abstract
In order to understand the potential target and related mechanism of action of Termina-lia Chebula treatment,network pharmacology and molecular docking methods were used in this experiment,and the challenge test of Salmonella from yak was performed.The active ingredients and potential targets of Terminalia Chebula were screened through HERB cluster identification database,TCMSP database and SwissTargetPrediction web page tool,and"gastroenteritis"was searched through OMIM and GeneCards database.Cytoscape and STRING databases were used to construct the Terminalia Chebula PPI network to screen out key targets,the intersection targets between Terminalia Chebula and enteritis were obtained through Venny platform,and gene ontol-ogy(GO)and Kyoto encyclopedia database of genes and genomics(KEGG)were enriched through DAVID database.The core target of screening was verified by molecular docking.After that,the gastrointestinal inflammation model of mice was established,the pathological changes of gastroin-testinal tract were observed,and the effect of Terminalia Chebula on the target protein was veri-fied by Western blot test.The results showed that:after analyzing and sorting out 8 main active in-gredients of Terminalia Chebula,118 targets of Terminalia Chebula were screened,11 161 targets of gastroenteritis and 100 targets of intersection were obtained;the core targets of PTGS2,CASP3,SLC3A2,Bax,Bcl-2 and TP53 of Terminalia Chebula and enteritis were obtained through PPI network.GO and KEGG enrichment analysis collected 337 items and 138 items,respectively,mainly related to chemokine pathway,PI3K-Akt signaling pathway,apoptosis related pathway,i-ron ion transport related pathway,NF-κB signaling pathway,etc.The results of molecular docking showed that chebulidic acid,the first active component of chebulidic acid,can bind to Bax,Bcl-2,PTGS2 and SLC3A2 through hydrogen bonding,hydrophobic action,π-π packing force and other intermolecular forces.The pathological tissue sections showed that Terminalia Chebula could sig-nificantly recover gastrointestinal tissue injury.Western blot test results showed that Terminalia Chebula can regulate the process of apoptosis and iron death through Bax/Bcl-2 and PTGS2/SLC3A2 pathways to achieve the effect of treating intestinal inflammatory damage.The results showed that Terminalia Chebula can regulate the occurrence and development of enteritis by regu-lating apoptosis and iron death through Bax/Bcl-2 and PTGS2/SLC3A2 pathways.Terminalia Chebula has the characteristics of multi-target and multi-pathway in the treatment of enteritis.
基金项目
现代农业产业技术体系资助项目(CARS-37)
西藏自治区科技厅2019年度重点基金资助项目(XZ201902NB04)
西藏农牧学院研究生创新计划基金资助项目(YJS2022-17)
维普
万方数据