梁霄;张筠昊;白皓天;李娅兰;孙淑慧;张倩倩;杨婧;王锐;
LIANG Xiao;ZHANG Jun-hao;BAI Hao-tian;LI Ya-lan;SUN Shu-hui;ZHANG Qian-qian;YANG Jing;WANG Rui;Heilongjiang University of Traditional Chinese Medicine;Basic Medical College of Heilongjiang University of Traditional Chinese Medicine;

• 1:黑龙江中医药大学
• 2:黑龙江中医药大学基础医学院

摘要(Abstract)：

利用网络药理学、分子对接技术和体外细胞实验，研究乌药抗胃癌的主要活性成分和潜在作用机制。利用中药系统药理学分析平台(TCMSP)数据库、在线人类孟德尔遗传(OMIM)和GeneCards数据库提取乌药活性成分，并预测了乌药与胃癌有关靶点。筛选二者共同的潜在功能靶位之后，再利用STRING数据库实现与二者共同靶点的蛋白相互作用(PPI)网络构建。利用DAVID数据库进行GO和KEGG富集分析；以STRING和DAVID数据库为基础，运用Cytoscape 3.7.2软件构建“活性成分-靶点”网络和“活性成分-靶点-通路”网络；利用AutoDock Vina进行分子对接，预测有效成分与关键作用靶点的结合度，最后对关键靶点和通路进行体外实验验证。预测结果共获得乌药活性成分9个、相关靶点179个，乌药与胃癌的共同靶点107个，靶点共涉及生物过程693种，细胞组成57种，分子功能129种，参与肿瘤抗原p53、缺氧诱导因子1等信号通路161条；分子对接结果显示核心成分吉马酮与TP53有较高的结合活性。最后体外实验将筛选出的乌药活性成分吉马酮对人胃癌细胞SGC-7901进行核心靶点和通路的初步验证，结果表明吉马酮可明显抑制胃癌细胞增殖并通过调控p53、Bax、Bcl-2等关键蛋白的表达来诱导SGC-7901凋亡。综上所述，乌药通过多成分、多靶点、多通路协同作用控制胃癌的发生发展，为今后临床进一步探讨乌药治疗胃癌的机制提供理论依据。
The present study explored the main active ingredients and the underlying mechanism of Linderae Radix the treatment of gastric cancer by network pharmacology, molecular docking, and in vitro cell experiments. TCMSP, OMIM and GeneCards database were used to obtain the active ingredients of Linderae Radix to predict the related targets of both Linderae Radix and gastric cancer. After screening the common potential action targets, the STRING database was used to construct the PPI network for protein interaction of the two common targets. Enrichment analysis of GO and KEGG by DAVID database. Based on STRING and DAVID platform data, Cytoscape software was used to construct an "active ingredient-target" network and an "active ingredient-target-pathway" network. Molecular docking was performed using the AutoDock Vina to predict the binding of the active components to the key action targets, and finally the key targets and pathways were verified in vitro. According to the prediction results, there were 9 active components, 179 related targets of Radix Linderae, 107 common targets of Linderae Radix and gastric cancer, 693 biological processes, 57 cell compositions, and 129 molecular functions involved in the targets, and 161 signaling pathways involved in tumor antigen p53, hypoxia-indu-cible factor 1, etc. Molecular docking results showed that the core component, jimadone, had high binding activity with TP53. Finally, in an in vitro experiment, the screened radix linderae active ingredient gemmadone is used for preliminarily verifying the core targets and pathways of the human gastric cancer cell SGC-7901, The results showed that germacrone could significantly inhibit the proliferation of gastric cancer cells and induce the apoptosis of SGC-7901 by regulating the expression of p53, Bax, Bcl-2 and other key proteins. In summary, Radix Linderae can control the occurrence and development of gastric cancer through multi-components, multi-targets and multi-pathways, which will provide theoretical basis for further clinical discussion on the mechanism of Radix Linderae in treating gastric cancer.

关键词(KeyWords)： 乌药;胃癌;网络药理学;分子机制;实验验证
Linderae Radix;gastric cancer;network pharmacology;molecular mechanism;experimental validation

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(81603418,82074271);; 黑龙江优秀青年人才项目(2020YQ05)

作者(Authors): 梁霄;张筠昊;白皓天;李娅兰;孙淑慧;张倩倩;杨婧;王锐;
LIANG Xiao;ZHANG Jun-hao;BAI Hao-tian;LI Ya-lan;SUN Shu-hui;ZHANG Qian-qian;YANG Jing;WANG Rui;Heilongjiang University of Traditional Chinese Medicine;Basic Medical College of Heilongjiang University of Traditional Chinese Medicine;

DOI: 10.19540/j.cnki.cjcmm.20220529.501

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