广西医学

目的　基于网络药理学及生物信息学方法探讨壮方消积化肝方通过调控铁死亡治疗肝癌的作用机制。方法　通过中药系统药理学数据库与分析平台筛选消积化肝方的活性成分及其作用靶点；通过DisGeNET、GeneCards®、OMIM®数据库筛选肝癌相关靶点；通过GeneCards®、FerrDb数据库筛选铁死亡相关靶点。将上述三类靶点的交集靶点导入STRING数据库构建蛋白⁃蛋白相互作用网络，并利用Cytoscape 3.8.0软件基于拓扑参数对交集靶点进行排序。通过Metascape数据库对交集靶点进行功能及通路富集分析。利用Cytoscape 3.8.0软件构建“消积化肝方活性成分⁃交集靶点⁃KEGG信号通路”网络，通过度值筛选核心活性成分。利用GEPIA2数据库分析交集靶点在正常肝组织与肝癌组织中的表达差异，以筛选核心靶点。使用AutoDockTools 1.5.6软件对核心活性成分及核心靶点对应蛋白进行分子对接分析。结果　获得消积化肝方活性成分的作用靶点197个、肝癌相关靶点1 578个、铁死亡相关靶点692个，取交集后获得交集靶点36个。交集靶点参与的生物学过程主要有程序性细胞死亡的正向调控、凋亡信号通路的调控等，富集的细胞组分主要有膜筏、膜微区等，涉及的分子功能主要有RNA聚合酶Ⅱ特异性DNA结合转录因子结合、DNA结合转录因子结合等；交集靶点主要富集的信号通路包括癌症相关通路、白细胞介素17信号通路、丝裂原活化蛋白激酶信号通路、缺氧诱导因子1信号通路等。筛选得到槲皮素、山柰酚、异鼠李素、异泽兰黄素4个核心活性成分，PARP1、CAV1、CDKN2A、HSPB1、NQO1 5个核心靶点。分子对接分析结果提示，核心活性成分与核心靶点对应蛋白具有较强的结合力。结论　壮方消积化肝方中的槲皮素、山柰酚、异鼠李素、异泽兰黄素等核心活性成分，可能通过作用于PARP1、CAV1、CDKN2A、HSPB1、NQO1 等基因来调控铁死亡，从而发挥治疗肝癌的作用。

Objective To explore the mechanism of Zhuangfang Xiaoji Huagan Prescription for the treatment of liver cancer by regulating ferroptosis based on network pharmacology and bioinformatics. Methods Active components and their effect targets of Xiaoji Huagan Prescription were screened by the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform. Targets related to liver cancer were screened by the databases of DisGeNET, GeneCards®, and OMIM®. Targets related to ferroptosis were screened by the databases of GeneCards® and FerrDb. The intersection targets of the aforementioned 3 categories were imported into the STRING database for establishing protein⁃protein interaction network, and the Cytoscape 3.8.0 software was used to rank the intersection targets based on topological parameters. Functional and pathway enrichment analyses on the intersection targets were conducted by the Metascape database. The “active components of Xiaoji Huagan Prescription⁃intersection targets⁃KEGG signaling pathway” network was established by employing the Cytoscape 3.8.0 software, and the core active components were screened by degree value. The differential expression between the intersection targets in normal liver tissues and liver cancerous tissues was analyzed by using the GEPIA2 database for screening the core targets. Molecular docking analysis of the core active components and core targets corresponding proteins was conducted by using the AutoDockTools 1.5.6 software. Results A total of 197 effect targets, 1578 targets related to liver cancer, 692 targets related to ferroptosis of active components of Xiaoji Huagan Prescription were obtained, and a total of 36 intersection targets after obtaining the intersection. The biological processes involved in the intersection targets mainly contained positive regulation of programmed cell death and regulation of apoptosis signaling pathway, etc. The cellular compositions enriched were mainly membrane rafts and membrane microregions, etc. The molecular functions involved mainly included RNA polymerase Ⅱ specific DNA⁃binding transcription factor binding and DNA⁃binding transcription factor binding, etc. The signaling pathways enriched in the intersection targets mainly contained cancer⁃related pathway, interleukin 17 signaling pathway, mitogen⁃activated protein kinase signaling pathway, hypoxia inducible factor 1 signaling pathway, etc. Four core active components, namely, quercetin, kaempferol, isorhamnetin, and eupatilin, and five core targets (PARP1, CAV1, CDKN2A, HSPB1, and NQO1) were screened. The results of molecular docking analysis suggested that the core active components had strong binding capacity with the corresponding proteins of the core targets. Conclusion The core active components with respect to quercetin, kaempferol, isorhamnetin, and eupatilin in Zhuangfang Xiaoji Huagan Prescription can regulate ferroptosis, so as to exert effect for the treatment of liver cancer through acting on PARP1, CAV1, CDKN2A, HSPB1, and NQO1 genes, etc.

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