基于网络药理学和分子对接探讨香砂六君子汤治疗幽门螺杆菌相关性胃炎的作用机制

doi:10.3976/j.issn.1002-4026.2023.04.007

摘要/Abstract

摘要：

利用网络药理学和分子对接方法分析香砂六君子汤的有效活性成分及其治疗幽门螺杆菌相关性胃炎的分子机制。通过中药系统药理学数据库与分析平台、GeneCards数据库和OMIM数据库筛选香砂六君子汤的药物活性化合物、药物作用靶点基因及幽门螺杆菌相关性胃炎疾病相关靶点,并对药物作用靶点和疾病相关靶点进行维恩分析;应用Cytoscape软件以及STRING数据库分别构建药物-化合物-潜在靶点相互作用网络及蛋白质-蛋白质相互作用网络图。使用DAVID数据库对交集靶点进行基因本体功能和京都基因与基因组百科全书通路富集分析。通过AutoDock PyMOL等软件对关键成分和作用靶点进行分子对接。通过吉姆萨染色法和CCK-8法测定细胞凋亡率,WesternBlot检测相关靶蛋白表达。最终筛选出香砂六君子汤方中槲皮素、木犀草素和山柰酚等122种药物活性化合物,可能通过作用于STAT3、TP53、AKT1等101个潜在靶点以及Toll样受体信号通路、TNF信号通路、T细胞受体信号通路等109条通路参与螺杆菌相关性胃炎的治疗。分子对接显示药物活性成分槲皮素、β-谷甾醇、木犀草素与靶点蛋白STAT3、TP53、AKT1有较好的亲和力。与模型组比较,用香砂六君子汤处理后,GES-1细胞的细胞核深染加深,细胞凋亡率显著下降,p-STAT3蛋白表达也显著降低。研究认为香砂六君子汤治疗幽门螺杆菌相关性胃炎以多成分、多靶点、多途径发挥其抑菌、抗炎的作用。

关键词: 香砂六君子汤, 幽门螺杆菌相关性胃炎, 网络药理学, 分子对接, 作用机制

Abstract:

This study aimed to analyze the active ingredients of Xiangsha Liujunzi Decoction and its molecular mechanism in treating Helicobacter pylori-associated gastritis using network pharmacology and molecular docking. The drug active compounds, drug target genes, and disease-related targets of H. pylori-associated gastritis in Xiangsha Liujunzi Decoction were screened using Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform, GeneCards database, and OMIM database, and the drug targets and disease-related targets were analyzed using Venn analysis. Cytoscape software and STRING database were used to construct drug-compound potential target interaction network and protein-protein interaction network, respectively. Gene Ontology function and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis were performed for intersection targets using the DAVID database. The key components and targets were docked using AutoDock PyMOL and other software. The apoptosis rate was determined with Jimsa staining and CCK-8 assay, and the expressions of the related target proteins were detected with western blot. Finally, 122 active compounds, such as quercetin, luteolin, and kaempferol, in Xiangsha Liujunzi Decoction were screened out. These genes may be involved in the treatment of H. pylori-associated gastritis by acting on 101 potential targets, such as STAT3, TP53, and AKT1, as well as 109 pathways, such as toll-like receptor, TNF, and T-cell receptor signaling pathways. Molecular docking showed that quercetin, β-sitosterol, and luteolin had good affinity for the target proteins STAT3, TP53, and AKT1. Compared with the model group, after treatment with Xiangsha Liujunzi Decoction, the nuclear hyperchromism of GES-1 cells was enhanced, the apoptosis rate was significantly decreased, and the expression of p-STAT3 was also significantly decreased. Based on these findings, it can be concluded that Xiangsha Liujunzi Decoction exerts antibacterial and anti-inflammatory effects in the treatment of H. pylori-associated gastritis in multiple ways via multiple components and targets.

Key words: Xiangsha Liujunzi Decoction, Helicobacter pylori-associated gastritis, network pharmacology, molecular docking, mechanism of action

中图分类号:

R285

尹志鹏, 高云云, 刘文文, 郭蓬勃, 赵英会. 基于网络药理学和分子对接探讨香砂六君子汤治疗幽门螺杆菌相关性胃炎的作用机制[J]. 山东科学, 2023, 36(4): 52-60.

YIN Zhipeng, GAO Yunyun, LIU Wenwen, GUO Pengbo, ZHAO Yinghui. Exploring the mechanism of Xiangsha Liujunzi Decoction in treating Helicobacter pylori-associated gastritis based on network pharmacology and molecular docking technology[J]. Shandong Science, 2023, 36(4): 52-60.

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药物	MOL ID	共有活性成分
党参、半夏、砂仁、木香	MOL000449	豆甾醇(stigmasterol)
党参、甘草	MOL003896	7-甲氧基-2-甲基异黄酮(7-methoxy-2-methyl isoflavone)
半夏、砂仁	MOL001755	24-乙基胆甾-4-烯-3-酮(24-ethylcholest-4-en-3-one)
半夏、砂仁	MOL000358	β-谷甾醇(beta-Sitosterol)
陈皮、甘草、木香	MOL000359	谷甾醇(sitosterol)
陈皮、甘草	MOL004328	柚皮素(naringenin)
甘草、木香	MOL000211	丁子香萜(mairin)

核心靶点	有效活性成分	结合自由能/(kJ·mol^-1)
	槲皮素(quercetin)	8.8
STAT3	β-谷甾醇(Beta-Sitosterol)	8.2
	木犀草素(Luteolin)	8.8
	槲皮素(quercetin)	8.9
TP53	β-谷甾醇(Beta-Sitosterol)	7.6
	木犀草素(Luteolin)	8.9
	槲皮素(quercetin)	9.1
AKT1	β-谷甾醇(Beta-Sitosterol)	6.9
	木犀草素(Luteolin)	7.5