目的 基于網(wǎng)絡(luò)藥理學(xué)和分子對接研究探討大蒜Allii Sativi Bulbus抗幽門螺桿菌(Hp)感染的活性成分及可能的作用機(jī)制,并通過體外抑菌試驗(yàn)驗(yàn)證大蒜素對Hp菌株的抗菌活性。方法 利用HERB數(shù)據(jù)庫獲取大蒜的活性成分,并通過PharmMapper、Swiss Target Prediction、BATMAN-TCM數(shù)據(jù)庫獲取活性成分靶點(diǎn)并構(gòu)建"藥物-成分-靶點(diǎn)"網(wǎng)絡(luò),篩選出主要活性成分。通過GeneCards、OMIM、DRUGBANK和DisGeNET數(shù)據(jù)庫收集與Hp感染相關(guān)的靶點(diǎn),取藥物與疾病靶點(diǎn)交集,篩選出潛在靶點(diǎn)。利用STRING數(shù)據(jù)庫獲取潛在靶點(diǎn)的互作關(guān)系,并在Cytoscape中構(gòu)建蛋白相互作用(PPI)網(wǎng)絡(luò),篩選出核心靶點(diǎn)。在DAVID數(shù)據(jù)庫中進(jìn)行潛在靶點(diǎn)的基因本體論(GO)生物分析與京都基因與基因組百科全書(KEGG)通路富集分析。通過Autodock進(jìn)行活性成分與核心靶點(diǎn)的分子對接。用體外抗菌試驗(yàn)檢測大蒜素的抗Hp活性。結(jié)果 共篩選出36個(gè)大蒜活性成分,291個(gè)潛在靶點(diǎn),PPI網(wǎng)絡(luò)分析得到腫瘤蛋白p53(TP53)、酪氨酸蛋白激酶(SRC)、信號傳導(dǎo)和轉(zhuǎn)錄激活蛋白3(STAT3)、熱休克蛋白90α家族A類成員1(HSP90AA1)、絲裂原激活的蛋白激酶(MAPK3)5個(gè)核心靶點(diǎn)。GO分析得到954個(gè)生物過程,KEGG分析得到176條通路。分子對接結(jié)果顯示,大蒜活性成分均與核心靶點(diǎn)有較好的結(jié)合能力。體外抑菌試驗(yàn)顯示大蒜素對3株Hp菌均有抑菌活性,最小抑菌濃度為2 mg/mL。結(jié)論 大蒜中的活性成分可能通過作用于TP53、SRC、STAT3、HSP90AA1、MAPK3 5個(gè)核心靶點(diǎn)參與Hp感染相關(guān)過程,其機(jī)制與磷脂酰肌醇3-激酶(PI3K)-蛋白激酶B(Akt)、叉頭框蛋白O(FoxO)、低氧誘導(dǎo)因子-1(HIF-1)等信號通路有關(guān),且大蒜素對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

新疆維吾爾自治區(qū)科技項(xiàng)目計(jì)劃(2021A03002)