基于网络药理学分析健脾通络利水方治疗肝郁脾虚兼血瘀型肝硬化失代偿期作用机制
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R285

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国家自然科学基金项目(81774453);河南省中医药科学研究专项项目(2023ZYZD13,2022ZY1147,2023ZY2139);河南省第三批中医药学科拔尖人才项目(豫卫中医药科教函〔2025〕15号);河南省医学科技攻关计划联合共建项目(LHGJ20250243)


Analysis of Mechanism of Action of Jianpi Tongluo Lishui Prescription in Treating Decompensated Cirrhosis of Liver-Depression and Spleen-Deficiency with Blood Stasis Type Based on Network Pharmacology
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    摘要:

    目的:利用网络药理学及分子对接技术分析健脾通络利水方治疗肝郁脾虚兼血瘀型肝硬化失代偿 期(DLC) 的作用机制。方法:采用中药系统药理学数据库与分析平台(TCMSP)、本草组鉴(HERB) 搜索健 脾通络利水方的活性成分、作用靶点,将Uniprot数据库标准化后的靶点取并集,得到该复方对应数据(药物- 成分-靶点),利用Cytoscape 软件构建药物成分相互作用网络。在Genecards、美国国家生物技术信息中 心(NCBI)、在线人类孟德尔遗传(OMIM) 数据库中检索DLC的对应靶点。根据肝郁脾虚兼血瘀型DLC的临 床主要表型在中医证候本体及多维定量关联计算平台(SoFDA) 检索相关靶点。疾病、证候靶点取交集,得到 病证对应的靶点。将方药成分与病证的共有基因靶点上传至STRING数据库进行蛋白质互作(PPI) 分析并筛 选关键靶点。通过Metascape在线分析系统进行基因本体(GO) 功能和京都基因与基因组百科全书(KEGG) 通路富集分析。并对关键成分及核心靶点进行分子对接验证。结果:健脾通络利水方16味核心药物共含活性 成分127个,相关靶点443个。获得肝郁脾虚兼血瘀型DLC疾病靶点921个,通过SOFDA平台检索的主症及次 症对应疾病靶点共156个。健脾通络利水方、DLC以及肝郁脾虚兼血瘀证的交集靶点共19个。健脾通络利水方 前3 个核心成分为槲皮素、β-谷甾醇、叶黄素。PPI 网络分析获得白细胞介素-6 (IL-6)、肿瘤蛋白 p53(TP53)、糖皮质激素受体(NR3C1)、骨髓细胞瘤癌基因(MYC)、表皮生长因子受体(EGFR)、转化生 长因子β1(TGFβ1) 等核心靶点。富集分析显示健脾通络利水方的16味核心药物主要在受体复合物、膜筏、 膜微区等区域发挥蛋白激酶调节剂活性、激酶调节剂活性、转录因子结合等生物过程。分子对接显示槲皮素、 β-谷甾醇、叶黄素活性成分与NR3C1、MYC结合稳定。结论:健脾通络利水方可能通过核心成分槲皮素、 β-谷甾醇、叶黄素等作用于NR3C1、MYC 等核心靶点,调节叉头框蛋白O (FoxO) 等信号通路,干预对 miRNA转录的积极调控等生物学进程,发挥治疗肝郁脾虚兼血瘀型DLC的作用。

    Abstract:

    Abstract: Objective: To analyze the mechanism of action of Jianpi Tongluo Lishui Prescription in treating decompensated cirrhosis (DLC) of liver-depression and spleen-deficiency with blood stasis type using network pharmacology and molecular docking technology. Methods:The active ingredients and corresponding targets of Jianpi Tongluo Lishui Prescription were retrieved from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform( TCMSP) and the HERB database. The targets standardized by the UniProt database were combined to obtain the "medicine-ingredient-target" data of the compound, and the medicine-ingredient interaction network was constructed using Cytoscape software. The targets related to DLC were searched in the GeneCards,National Center for Biotechnology Information (NCBI), and Online Mendelian Inheritance in Man (OMIM) databases. According to the main clinical phenotypes of DLC of liver-depression and spleen-deficiency with blood stasis type,relevant targets were retrieved from the Syndrome Ontology for Traditional Chinese Medicine and Multidimensional Quantitative Association Calculation Platform (SoFDA). The intersection of disease targets and syndrome targets was taken to obtain the diseasesyndrome corresponding targets. The common gene targets of the prescription′s ingredients and the "disease-syndrome" were uploaded to the STRING database for protein-protein interaction (PPI) analysis,and key targets were screened. The Metascape online analysis system was used for Gene Ontology (GO) functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. Molecular docking verification was performed on key ingredients and core targets. Results:A total of 127 active ingredients and 443 related targets were identified from the 16 core drugs of Jianpi Tongluo Lishui Prescription. A total of 921 disease targets of DLC of liverdepression and spleen-deficiency with blood stasis type were obtained, and 156 disease targets corresponding to primary and secondary symptoms were retrieved via the SoFDA platform. There were 19 overlapping targets among Jianpi Tongluo Lishui Prescription, DLC, and liver-depression and spleen-deficiency with blood stasis syndrome. The top three core ingredients of the prescription were quercetin, β-sitosterol, and luteolin. PPI network analysis identified core targets including interleukin-6 (IL-6),tumor protein p53 (TP53),glucocorticoid receptor (NR3C1),V-myc avian myelocytomatosis viral oncogene homolog (MYC),epidermal growth factor receptor (EGFR),and transforming growth factor beta 1 (TGFβ1). Enrichment analysis showed that the 16 core Chinese herbs of the prescription mainly exerted effects in regions such as receptor complexes, membrane rafts, and membrane microdomains, involving biological processes including protein kinase modulator activity, kinase modulator activity, and transcription factor binding. Molecular docking results demonstrated stable binding between the active ingredients (quercetin,β-sitosterol, and luteolin) and the core targets (NR3C1,and MYC). Conclusion:Jianpi Tongluo Lishui Prescription may act on core targets such as NR3C1 and MYC through its core ingredients (quercetin,β-sitosterol,and luteolin) to regulate signaling pathways like FoxO and biological processes such as the positive regulation of miRNA transcription,thereby exerting a therapeutic effect on DLC of liver-depression and spleen-deficiency with blood stasis type.

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娄静,潘晔,邢崇溢,崔伟锋,赵移畛,侯留法.基于网络药理学分析健脾通络利水方治疗肝郁脾虚兼血瘀型肝硬化失代偿期作用机制[J].新中医,2025,57(23):219-226

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