基于网络药理学和分子对接探究芹菜素缓解奶牛热应激及低氧应激的潜力与机制  

The Potential and Mechanisms of Apigenin to Relieve Heat Stress and Hypoxia in Dairy Cows Based on Network Pharmacology and Molecular Docking

作　　者：刘卓琳 刘红云[1] LIU ZhuoLin;LIU HongYun(College of Animal Science,Zhejiang University,The Institute of Dairy Science,Hangzhou 310058)

机构地区：[1]浙江大学动物科学学院,奶业科学研究所,杭州310058

出　　处：《中国农业科学》2024年第5期1010-1022,共13页Scientia Agricultura Sinica

基　　金：国家自然科学基金(32072756);国家奶牛产业技术体系(CARS-36)。

摘　　要：【目的】利用网络药理学和分子对接方法,系统预测芹菜素缓解奶牛热应激及低氧应激的作用机制。为芹菜素的应用提供参考。【方法】首先从TCMSP(中药系统药理学数据库)和Gene Cards(基因数据库)分别检索“Apigenin”“heat stress”“hypoxia”,获得芹菜素、热应激和低氧应激相关靶点,韦恩交集取得芹菜素缓解双重应激(热应激+低氧)的靶点集。利用STRING数据库构建双重靶点蛋白-蛋白互作(PPI)网络,使用Cytoscape 3.9.1软件的Cyto NCA插件分别对网络节点中心性进行计算,筛选得到核心靶点。利用David数据库的分析工具“Functional Annotation”对度值排名前30的芹菜素缓解双重应激靶点进行基因本体(GO)功能和京都基因与基因组百科全书(KEGG)通路富集分析。同时,对以上靶点进行分子复合物检测分析(MCODE)得到核心基因簇,并进一步对核心基因簇进行GO和KEGG分析。根据KEGG通路富集结果与核心靶点信息构建芹菜素-双重应激靶点-信号通路网络。最后,使用Autodock vina程序将芹菜素与核心靶点逐一进行分子对接,利用Pymol软件和PLIP网站分析其相互作用模式并将结果可视化。【结果】TCMSP数据库检索得到芹菜素靶点68个,Gene Cards数据库中获得低氧相关基因6 661个,热应激相关基因9 046个,获得三者的交集靶点56个,即芹菜素缓解双重应激的可能靶点。靶点PPI网络的节点数为56,边数为436,平均节点度值为15.6,平均局部聚类系数为0.728,PPI富集P值<1.0×10^(-16)。核心靶点分别为AKT1(RAC-α丝氨酸/苏氨酸蛋白激酶)、TP53(细胞肿瘤抗原p53)、TNF(肿瘤坏死因子)、CASP3(细胞凋亡蛋白酶3)、INS(胰岛素)、BCL2L1(BCL-2样蛋白1)、VEGFA(血管内皮生长因子A)、HIF1A(缺氧诱导因子1-α)、PTGS2(前列腺素G/H合酶2)和SREPINE1(纤溶酶原激活物抑制剂1),分子对接结果显示芹菜素与上述核心靶点均可稳定结合。GO功能富集分析(P<0.0【Objective】The study aimed to predict the mechanisms of apigenin in relieving heat stress and hypoxia stress in dairy cows by using network pharmacology and molecular docking so as to provide a reference for the application of apigenin.【Method】Firstly,the targets related to apigenin,heat stress,and hypoxia stress were obtained from TCMSP and GeneCards databases,and apigenin targets in relieving dual stress were obtained by Venn intersection.The protein-protein interaction(PPI)network was constructed using the STRING database,and the core targets were identified by analyzing the network node centrality using the CytoNCA plugin in Cytoscape 3.9.1 software.Autodock vina program was used to perform molecular docking of apigenin with the identified targets.Perform gene ontology(GO)functional and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis on the top 30 ranked targets based on degree centrality using Functional Annotation,the analysis tool of David database.Additionally,Molecular complex detection analysis(MCODE)of the above targets yielded the core gene clusters,then GO and KEGG analysis were performed.Apigenin-dual stress targets-signaling pathway network model were constructed through the KEGG pathway enrichment results and core target information.Finally,apigenin was docked to the core targets using the Autodock vina program,and the results were visualized using the Pymol software and the PLIP website.【Result】The TCMSP database retrieved 68 targets of apigenin,6661 hypoxia-related genes and 9046 heat stress-related genes were obtained in GeneCards database,and 56 apigenin targets in relieving dual stress were obtained.The number of nodes in the target PPI network was 56,the number of edges was 436,the average node degree value was 15.6,the mean local clustering coefficient was 0.728,and PPI enrichment P value was less than 1.0×10^(-16).The core targets were AKT1(RAC-alpha serine/threonine-protein kinase),TP53(Cellular tumor antigen p53),TNF(Tumor necrosis factor),CASP3(Casp

关 键 词：芹菜素 奶牛 热应激 低氧 网络药理学 分子对接 

分 类 号：S853.74[农业科学—临床兽医学]