基于网络药理学研究参附注射液治疗脓毒症的作用机制

Mechanism of Shenfu Injection in the treatment of sepsis basedon network pharmacology

利用网络药理学和分子对接方法研究参附注射液治疗脓毒症的物质基础及其可能的作用机制。通过中药系统药理学数据库与分析平台和SwissTargetPrediction数据库得到参附注射液活性成分及其药物靶点,通过GeneCards、OMIM和Drugbank数据库筛选得到脓毒症相关靶点,STRING数据库和Cytoscape软件构建蛋白质-蛋白质相互作用网络;使用Metascape平台进行基因本体(gene ontology,GO)和基于京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)通路分析;应用Cytoscape软件构建成分-靶点-通路网络;应用AutoDock软件对关键成分和靶点进行分子对接;尾静脉注射LPS建立小鼠脓毒症模型,利用ELISA和Western blot验证参附注射液对脓毒症的治疗效果。结果表明:参附注射液的潜在活性成分共有34个,其治疗脓毒症的关键靶点137个,GO和KEGG分析靶点主要涉及炎症、细胞凋亡和信号转导等生物学过程,通过PI3K-Akt和MAPK等信号通路治疗脓毒症;分子对接结果显示关键成分与对应靶点具有较好的结合活性;体内实验验证结果表明,参附注射液能够抑制脓毒症小鼠血清中的炎症因子TNF-α和IL-6表达;Western blot结果表明,参附注射液可以上调肝组织中ERK表达(P<0.01),下调p-ERK的表达(P<0.01)。该研究发现参附注射液是通过多成分-多靶点-多通路的作用方式发挥抗脓毒症作用,为明确其抗脓毒症的作用机制提供了科学依据。

This study aimed to study the material basis of Shenfu Injection in the treatment of sepsis and its possible mechanism of action using network pharmacology and molecular docking methods.Traditional Chinese Medicine System Pharmacology Database and Analysis Platform and Swiss Target Prediction database were used to identify the potential active ingredients and drug targets of Shenfu Injection.Screening of disease targets was performed using GeneCards,OMIM,and Drugbank databases.String database and Cytoscape software were used to construct a protein-protein interaction network.Metascape platform was used for gene ontology(GO)enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analyses.Cytoscape software was used to construct a component-target-pathway network.Auto Dock software was used to perform molecular docking of the key components and targets.A mouse sepsis model was developed by injecting LPS into the tail vein.The therapeutic effect of Shenfu Injection on sepsis was verified using ELISA and western blot.The results revealed 34 potential active ingredients in Shenfu Injection and 137 key targets involved in sepsis treatment.The results of GO and KEGG analyses showed that these targets primarily involved biological processes such as inflammation,apoptosis,and signal transduction and played a role in sepsis treatment through signaling pathways such as PI3K-Akt and MAPK.The results of molecular docking revealed that the key components had good binding activities with the corresponding targets.In vivo experiments verified that Shenfu Injection inhibits the expression of inflammatory factors TNF-αand IL-6 in septic mice.Western blot results showed that Shenfu Injection could significantly upregulate ERK expression(P<0.01)and downregulate p-ERK expression(P<0.01)in liver tissues.This study found that Shenfu Injection exerts its antiseptic effect through a multicomponent-multitarget-multipathway mode of action,which provides an important scientific basis for clarifying its antisepsis mechanism.