目的 利用網(wǎng)絡(luò)藥理學(xué)探討百部發(fā)揮止咳作用的活性成分、作用靶點(diǎn)及相關(guān)通路,探究百部止咳的作用機(jī)制。方法 通過中藥系統(tǒng)藥理學(xué)數(shù)據(jù)庫(kù)和分析平臺(tái)(TCMSP)獲取并篩選百部主要活性成分及其對(duì)應(yīng)的作用靶點(diǎn),利用Uniprot蛋白質(zhì)數(shù)據(jù)庫(kù)將作用靶點(diǎn)標(biāo)準(zhǔn)化,利用Cytoscape構(gòu)建百部"成分-靶點(diǎn)"網(wǎng)絡(luò);利用GeneCards獲取并篩選疾病"咳嗽"的作用靶點(diǎn),利用VENNY獲取交集靶點(diǎn),將結(jié)果提交至String平臺(tái),獲取蛋白質(zhì)-蛋白質(zhì)相互作用(PPI)網(wǎng)絡(luò);使用metascape進(jìn)行基因本體(GO)富集分析與京都基因與基因組百科全書(KEGG)通路富集分析,并通過微生信平臺(tái)將結(jié)果進(jìn)行可視化,根據(jù)與咳嗽相關(guān)程度對(duì)通路進(jìn)行篩選,利用Cytoscape構(gòu)建百部"成分-靶點(diǎn)-通路"網(wǎng)絡(luò)。利用RT-PCR法檢測(cè)前列腺素內(nèi)過氧化物合成酶2(PTGS2)、胱天蛋白酶3(CASP3)、絲裂原活化蛋白激酶14(MAPK14)mRNA水平上的表達(dá)變化。結(jié)果 獲取及篩選得到百部活性成分28個(gè),作用靶點(diǎn)96個(gè),咳嗽靶點(diǎn)1170個(gè),交集靶點(diǎn)39個(gè);GO功能富集分析顯示其與對(duì)雌二醇、類固醇激素、脂多糖、藥物的反應(yīng)及血管形態(tài)發(fā)生及血管發(fā)育等相關(guān),KEGG通路富集分析顯示其與T細(xì)胞受體信號(hào)通路、Nod樣受體信號(hào)通路、MAPK信號(hào)通路、VEGF信號(hào)通路、NF-κB信號(hào)通路等相關(guān)。RT-PCR結(jié)果表明百部對(duì)PTGS2、CASP3、MAPK14表達(dá)有明顯的抑制作用。結(jié)論 百部可能通過抑制PTGS2、CASP3、MAPK14因子的表達(dá)發(fā)揮止咳作用,為探究百部止咳的作用機(jī)制提供了理論參考。

Objective To explore the active components, action targets and related pathways of Stemonae Radix in treatment of cough based on network pharmacology, and to explore the mechanism of relieving cough. Methods The main active components and their corresponding targets of Stemonae Radix were obtained and screened by TCMSP, and action targets were standardized using Uniprot protein database. Cytoscape was used to construct hundreds of “component-target” networks. GeneCards was used to obtain and screen the targets of “cough”, and VENNY was used to obtain the intersection targets, and the results were submitted to String platform to obtain the PPI network. Metascape was used for GO enrichment analysis and KEGG pathway enrichment analysis, and the results were visualized by micromessenger. Pathways were screened according to the degree of cough correlation, and Cytoscape was used to construct hundreds of “component - target – pathway” networks. Real-time PCR was used to detect the mRNA expression changes of PTGS2, CASP3 and MAPK14. Results 28 Active ingredients, 96 targets, 1 170 cough targets and 39 intersection targets were obtained. GO functional enrichment analysis showed that it was involved in the response to estradiol, steroid, lipopolysaccharide, drugs and related to vascular morphogenesis and vascular development. KEGG pathway enrichment analysis showed that it was related to T cell receptor signaling pathway, Nod-like receptor signaling pathway, MAPK signaling pathway, VEGF signaling pathway, NF-κB signaling pathway. Real-time PCR results showed that Stemonae Radix had significant inhibitory effect on PTGS2, CASP3 and MAPK14 mRNA expression. Conclusion Stemonae Radix may relieve cough by inhibiting the expression of PTGS2, CASP3 and MAPK14 mRNA, which provides theoretical reference for exploring the specific mechanism of Stemonae Radix in treatment of cough.

R285.5

國(guó)家自然科學(xué)基金面上項(xiàng)目（82174060）；黑龍江省自然科學(xué)基金資助項(xiàng)目（LH2021H074）；黑龍江省衛(wèi)生健康委科研課題（20210202040011）