网络药理学联合分子对接研究藏药六味木香丸治疗胃癌的分子作用机制

Network Pharmacology Combined with Molecular Docking to Study the Molecular Mechanism of Tibetan Medicine Liuwei Muxiang Pill in the Treatment of Gastric Cancer

目的通过网络药理学和分子对接的方法预测藏药六味木香丸治疗胃癌的主要活性成分和作用靶点。方法采用文献检索与TCMSP数据库收集藏药六味木香丸组成药材木香、豆蔻、余甘子、荜茇、巴夏嘎、余甘子的主要化学成分和作用靶点,从GeneCards、OMIM、PharmGkb、TTD、DrugBank五大疾病数据库共查找胃癌对应的靶点,将成分靶点和疾病靶点构建韦恩图。并通过GO和KEGG通路富集这些获得的交集靶点。在软件Cytoscape中构建“药物-活性化合物-靶点”网络,通过Cytoscape插件-CytoNCA筛选藏药六味木香丸治疗胃癌的核心化合物和核心靶点并绘制其关系网络。依据AutoDock Vina软件和Perl脚本对核心化合物与核心靶点进行分子对接。结果TCMSP数据库和文献查阅显示六味木香丸共有42个活性化合物映射301个靶点。从GeneCards、OMIM、PharmGkb、TTD、DrugBank数据库共获得胃癌相关疾病靶点8953个,成分靶点和疾病靶点取交集共获得271个交集靶点。GO功能富集分析表明主要涉及氧化胁迫响应、营养水平响应、细胞膜、DNA-结合转录激活活性、丝氨酸/苏氨酸激酶活性等过程。KEGG通路富集分析显示涉及PI3K/Akt、MAPK、AGE/RAGE等信号通路。分子对接发现槲皮素和木犀草素与MAPK1、JUN,山奈酚、β-谷甾醇与JUN,(-)-儿茶素-3-没食子酸与MAPK1、MAPK3、JUN、STAT3都有很好的结合能力。体外细胞实验发现六味木香丸能够有效抑制人胃癌细胞SGC-7901增殖,并显著抑制STAT3蛋白的表达。结论本研究证实六味木香丸具有多成分、多靶点、协同调节的特点,通过网络药理学预测了六味木香丸治疗胃癌的可能机制,为其活性成分研究和试验研究提供理论依据。

Objective To predict the main active ingredients and targets of Liuwei Muxiang(LWMX) pill in the treatment of gastric cancer based on network pharmacology combined with molecular docking.Methods The literature search and TCMSP database were used to collect the main chemical components and targets of LWMX Pill, which was composed of Radix Aucklandiae, Fructus Ammomi Rotundus, Phyllanthus emblica, Fructus Piperis Longi, Paxiaga, and Semen Punicae Granati. GO and KEGG pathway was used to analyze the intersection targets, and Cytoscape software was used to visualize an "active compound-target" network. The core compounds and core targets of LWMX Pill for the treatment of gastric cancer were deeply screened through the Network Analysis plug-in. In addition, AutoDock Vina software and Python script was used to construct the core active compound molecularly docked with the core target protein receptor.Results It showed that LWMX Pill had 42 active compounds mapped to 301 targets. A total of 8953 gastric cancer-related disease targets were obtained from GeneCards, OMIM, PharmGkb, TTD, and DrugBank databases, and a total of 271 intersection targets were obtained from the intersection of component targets and disease targets. GO analysis showed that it mainly involved processes such as oxidative stress response, nutrient level response,cell membrane, DNA-binding transcription activation, and serine or threonine kinase activity. KEGG pathway showed that PI3K/Akt, MAPK, AGE/RAGE and other signaling pathways were involved. Molecular docking found that quercetin and luteolin are match with MAPK1, JUN, kaempferol, β-sitosterol and JUN,(-)-catechin-3-gallic acid were able to bind the MAPK1, MAPK3, JUN, STAT3. In vitro cell experiments found that Liuwei Muxiang Pill can effectively inhibit the proliferation of human gastric cancer cells SGC-7901 and significantly inhibit the expression of STAT3 protein.Conclusion Tibetan formulae Liuwei Muxiang Pill has multi-component, multi-target and multi-path effects on gastric cancer.