目的 基于网络药理学和分子对接研究探讨大蒜Allii Sativi Bulbus抗幽门螺杆菌(Hp)感染的活性成分及可能的作用机制,并通过体外抑菌试验验证大蒜素对Hp菌株的抗菌活性。方法 利用HERB数据库获取大蒜的活性成分,并通过PharmMapper、Swiss Target Prediction、BATMAN-TCM数据库获取活性成分靶点并构建"药物-成分-靶点"网络,筛选出主要活性成分。通过GeneCards、OMIM、DRUGBANK和DisGeNET数据库收集与Hp感染相关的靶点,取药物与疾病靶点交集,筛选出潜在靶点。利用STRING数据库获取潜在靶点的互作关系,并在Cytoscape中构建蛋白相互作用(PPI)网络,筛选出核心靶点。在DAVID数据库中进行潜在靶点的基因本体论(GO)生物分析与京都基因与基因组百科全书(KEGG)通路富集分析。通过Autodock进行活性成分与核心靶点的分子对接。用体外抗菌试验检测大蒜素的抗Hp活性。结果 共筛选出36个大蒜活性成分,291个潜在靶点,PPI网络分析得到肿瘤蛋白p53(TP53)、酪氨酸蛋白激酶(SRC)、信号传导和转录激活蛋白3(STAT3)、热休克蛋白90α家族A类成员1(HSP90AA1)、丝裂原激活的蛋白激酶(MAPK3)5个核心靶点。GO分析得到954个生物过程,KEGG分析得到176条通路。分子对接结果显示,大蒜活性成分均与核心靶点有较好的结合能力。体外抑菌试验显示大蒜素对3株Hp菌均有抑菌活性,最小抑菌浓度为2 mg/mL。结论 大蒜中的活性成分可能通过作用于TP53、SRC、STAT3、HSP90AA1、MAPK3 5个核心靶点参与Hp感染相关过程,其机制与磷脂酰肌醇3-激酶(PI3K)-蛋白激酶B(Akt)、叉头框蛋白O(FoxO)、低氧诱导因子-1(HIF-1)等信号通路有关,且大蒜素对Hp菌有抑菌活性。

Objective To explore active components and mechanism of Allium sativum on inhibition of Hp infection based on network pharmacology and molecular docking, and verify the antibacterial activity of allicin on inhibition of Hp in vitro. Methods First, the active components of Allium sativum were obtained from HERB database. PharmMapper, Swiss Target Prediction, BATMAN-TCM database were used to obtain the active targets of components and the "medicine-component-target" network was constructed, screening out the main active components. Then targets related to Hp infection were collected from GeneCards, OMIM, DRUGBANK, and DisGeNET databases, and potential targets were selected by the common targets of active components and disease. Afterward, STRING database was used to obtain the interaction of targets, and Cytoscape was employed to establish the PPI network of potential targets, and screened for the core targets. The GO term enrichment analysis and KEGG pathway enrichment analysis were performed in DAVID database. Molecular docking between the active components and the key targets was performed by Autodock. Finally, the ntibacterial activity of allicin was verified by in vitro antibacterial test. Results There were thirty six Allium sativum active components and 291 potential targets were collected, and five core targets including TP53, SRC, STAT3, HSP90AA, and MAPK3 were identified by PPI network analysis. GO analysis resulted in 954 biological processes, and KEGG analysis resulted in 176 pathways. Molecular docking results showed that all the active components of Allium sativum had high binding ability with their core targets. In vitro antibacterial test showed the antibacterial activity of allicin on inhibition of Hp, with the minimum inhibitory concentration of 2.0 mg/mL. Conclusions Active components in Allium sativum may be involved in Hp infection by regulating five core targets, including TP53, SRC, STAT3, HSP90AA1, MAPK3, and the mechanisms are related to PI3K-Akt, FoxO, HIF-1 signaling pathways. Allicin show anti-Hp activity by in vitro antibacterial test.

R285

新疆维吾尔自治区科技项目计划(2021A03002)