当归四逆汤加吴茱萸生姜汤及其核心成分山柰酚对肝癌机制的研究

作者：梁 楠，李 峥，程紫轩，孙异强，牛杜升，黄 博

单位：山西医科大学附属山西省人民医院，山西 太原 030012

引用：引用：梁楠，李峥，程紫轩，孙异强，牛杜升，黄博.当归四逆汤加吴茱萸生姜汤及其核心成分山柰酚对肝癌机制的研究[J].中医药导报,2025,31(7):16-24.

DOI：10.13862/j.cn43-1446/r.2025.07.003

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摘要：目的：基于网络药理学、分子动力学模拟及免疫组化探究当归四逆汤加吴茱萸生姜汤（DSJWST）抗肝癌的作用机制。方法：借助中药系统药理学数据库与分析平台（TCMSP）对DSJWST所含8味中药的活性成分及潜在靶点进行筛选，整合GeneCards、OMIM、TCGA等数据库资源，获取与肝癌相关的靶点信息，再通过韦恩图分析，精准确定药物-疾病的交集基因。利用String数据库构建蛋白互作（PPI）网络，直观展示靶点间的相互作用关系，并采用Cytoscape软件的MCC算法识别DSJWST抗肝癌关键靶标，进一步筛选核心成分。通过基因本体论（GO）和京都基因和基因组百科全书（KEGG）对靶蛋白功能和相关通路进行富集分析，挖掘DSJWST抗肝癌的潜在分子机制。同时，借助Cytoscape构建“药物-核心成分-核心靶点”网络，呈现药物、成分与靶点之间的内在联系。采用Pymol插件和GROMACS 2016.4软件，开展分子对接及分子动力学模拟实验，从分子层面探究DSJWST活性成分与靶标的结合特性及稳定性。最后，通过免疫组化实验，对肝癌关键靶点表达及其与临床预后的相关性进行验证。结果：经筛选，共获得1 599个活性成分和4 124个肝癌相关靶点，确定了166个交集靶点。通过PPI网络筛选出10个关键靶标，包括IL6、TNF、STAT3、IL1B、AKT1、IFNG、TP53、NFKBIA、JUN和CASP3。KEGG富集分析显示，DSJWST治疗肝癌主要涉及AGE-RAGE信号通路、IL-17信号通路、TNF信号通路及HIF-1信号通路等。分子对接结果显示，DSJWST的10个活性组分与7个靶标的对接结合能均小于-5.0 kcal/mol，表明两者具有一定的结合活性。之后，选取结合能最高的山柰酚与CASP3进行分子动力学模拟，结果表明，在与山柰酚的结合过程中CASP3结构趋于稳定。免疫组织化学实验证实，CASP3在肝癌组织中呈低表达状态，且与良好预后相关。结论：DSJWST可能通过靶向CASP3，进而介导AGE-RAGE信号通路的激活，发挥抑制肝癌进展的作用。

关键词：肝癌；当归四逆汤加吴茱萸生姜汤；网络药理学；分子动力学模拟；CASP3

Abstract：

Objective: To investigate the mechanism of Danggui Sini Tang Jia Wuzhuyu Shengjiang Tang (DSJWST) in combating liver cancer based on network pharmacology, molecular dynamics simulation, and immunohistochemistry. Methods: The active components and potential targets of the eight herbs in DSJWST were screened using the traditional Chinese medicine systems pharmacology database and analysis platform (TCMSP). Liver cancer-related targets were obtained from the GeneCards, OMIM, and TCGA databases, and the drug-disease intersection genes were identified through a Venn diagram. The String database was used to construct a protein-protein interaction (PPI) network, and the MCC algorithm in Cytoscape software was employed to identify key targets of DSJWST against liver cancer and to screen core components. The functions and pathways of the target proteins were analyzed through gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) enrichment analyses. A "drug-core component-core target" network was constructed using Cytoscape. Molecular docking and molecular dynamics simulations were performed using the Pymol plugin and GROMACS 2016.4 software. Finally, immunohistochemical experiments were conducted to validate the expression of key liver cancer targets and their correlation with clinical prognosis. Results: A total of 1 599 active components and 4 124 liver cancer-related targets were screened, resulting in 166 intersection targets. Through the PPI network, 10 key targets were selected, including IL6, TNF, STAT3, IL1B, AKT1, IFNG, TP53, NFKBIA, JUN, and CASP3. KEGG enrichment analysis revealed that the treatment of liver cancer by DSJWST was primarily associated with the AGE-RAGE signaling pathway, IL-17 signaling pathway, TNF signaling pathway, and HIF-1 signaling pathway. Molecular docking results showed that the docking binding energies of the 10 active components of DSJWST and 7 targets were all below -5.0 kcal/mol, indicating significant binding activity. Subsequently, kaempferol, which exhibited the highest binding energy, was selected for molecular dynamics simulation with CASP3. The results demonstrated that the structure of CASP3 tended to stabilize during its binding process with kaempferol. Meanwhile, immunohistochemical experiments confirmed that CASP3 was lowly expressed in liver cancer tissues and that it was associated with a favorable prognosis. Conclusion: Danggui Sini Tang Jia Wuzhuyu Shengjiang Tang may inhibit the progression of liver cancer by targeting CASP3 and subsequently mediating the activation of AGE-RAGE signaling pathway.

Key words：liver cancer; Danggui Sini Tang Jia Wuzhuyu Shengjiang Tang; network pharmacology; molecular dynamics simulation; CASP3

发布时间：2026-01-06

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