参附汤治疗脓毒症的动物实验及网络药理学研究

Mechanism of Shenfu Decoction in the Treatment of Sepsis Based on Network Pharmacology and Experi-mental Verification

目的 基于网络药理学研究参附汤治疗脓毒症的物质基础及其可能的作用机制。方法 通过中药系统药理学分析平台(TCMSP)数据库查找参附汤组成药物人参、附子的活性成分和药物靶点;通过GeneCards、OMIM、DrugBank、PharmGkb及TTD数据库中获得脓毒症相关靶点,利用韦恩图得到药物靶点和疾病靶点的交集,得到参附汤治疗脓毒症的关键靶点;使用软件Cytoscape构建参附汤治疗脓毒症的“药物-活性成分-疾病-靶点”网络,并联合STRING平台构建关键靶点PPI网络;通过Bioconductor平台对关键靶点进行GO和KEGG富集分析。建立盲肠结扎穿孔诱导的脓毒症大鼠模型,进一步验证参附汤治疗脓毒症的作用机制。结果 参附汤共有45个活性化合物,作用于99个活性成分靶点,与1 564个脓毒症靶点取交集共得到46个作用靶点,包括Akt1、IL-1β、PPARG、PTGS2、JUN、CASP3等;GO功能富集分析和KEGG通路富集分析主要涉及炎症、细胞凋亡、营养、能量代谢、缺氧等,通过肿瘤坏死因子(TNF)信号通路、白细胞介素-17(IL-17)信号通路、细胞凋亡、NF-κB信号通路等发挥治疗脓毒症的作用。动物实验结果表明,参附汤有助于降低脓毒症大鼠血清的TNF-α和白细胞介素-6(IL-6)的表达水平,上调肠道组织中Akt磷酸化的表达水平(P <0.05)。结论 参附汤通过多成分-多靶点-多通路的作用方式治疗脓毒症,为其治疗脓毒症提供了一定的依据。

Objective:To study the material basis and possible mechanism of Shenfu Decoction(SFD)in the treatment of sepsis by network pharmacology.Methods:The active components and drug targets of ginseng root(Panax ginseng C.A.Mey,Renshen)and aconite root(Aconitum carmichaeli Debeaux,Fuzi)were searched through the database of Traditional Chinese Medicine System Pharmacology Analysis Platform(TCMSP).Screening of sepsis related targets was performed using GeneCards,OMIM,DrugBank,PharmGkb and TTD databases.Venn diagram was used to obtain the intersection of SFD targets and sepsis targets,and the key targets of SFD in the treatment of sepsis were obtained.Cytoscape software was used to construct a drug-component-target-pathway network.String database and Cytoscape software were used to construct a protein-protein interaction(PPI)network.Bioconductor platform was used for gene ontology(CO)enrichment and Kyoto Encyclopedia of Genesand Genomes(KECC)pathway analyses.To further verify the mechanism of SFD in the treatment of sepsis,a sepsis rat model induced by cecal ligation and perforation was established.The therapeutic effect of SFD on sepsis was verified using ELISA and Western bloting.Results:The results revealed 45 potential active ingredients in SFD and 99 key targets involved in sepsis treatment.A total of 46 target sites were obtained among 1564 sepsis targets,including Aktl,IL-1β,PPARG,PTGS2,JUN,CASP3,etc.GO enrichment analysis and KEGG pathway enrichment analysis mainly involved inflammation,apoptosis,nutrition,energy metabolism,hypoxia,etc.,which played a role in the treatment of sepsis through TNF signaling pathway,IL-17 signaling pathway,apoptosis,NF-kB signaling pathway,etc.The results of sepsis rats showed that SFD could reduce the expression levels of TNF-αand IL-6 in serum and up-regulate the phosphorylation level of Akt in intestinal tissue of septic rats(P<0.05).Conclusion:SFD plays a role in the treatment of sepsis through a multi-component,multi-target and multi-pathway mode of action,which has provided some basis for its treatment of sepsis.