目的 利用网络药理学探讨百部发挥止咳作用的活性成分、作用靶点及相关通路,探究百部止咳的作用机制。方法 通过中药系统药理学数据库和分析平台(TCMSP)获取并筛选百部主要活性成分及其对应的作用靶点,利用Uniprot蛋白质数据库将作用靶点标准化,利用Cytoscape构建百部"成分-靶点"网络;利用GeneCards获取并筛选疾病"咳嗽"的作用靶点,利用VENNY获取交集靶点,将结果提交至String平台,获取蛋白质-蛋白质相互作用(PPI)网络;使用metascape进行基因本体(GO)富集分析与京都基因与基因组百科全书(KEGG)通路富集分析,并通过微生信平台将结果进行可视化,根据与咳嗽相关程度对通路进行筛选,利用Cytoscape构建百部"成分-靶点-通路"网络。利用RT-PCR法检测前列腺素内过氧化物合成酶2(PTGS2)、胱天蛋白酶3(CASP3)、丝裂原活化蛋白激酶14(MAPK14)mRNA水平上的表达变化。结果 获取及筛选得到百部活性成分28个,作用靶点96个,咳嗽靶点1170个,交集靶点39个;GO功能富集分析显示其与对雌二醇、类固醇激素、脂多糖、药物的反应及血管形态发生及血管发育等相关,KEGG通路富集分析显示其与T细胞受体信号通路、Nod样受体信号通路、MAPK信号通路、VEGF信号通路、NF-κB信号通路等相关。RT-PCR结果表明百部对PTGS2、CASP3、MAPK14表达有明显的抑制作用。结论 百部可能通过抑制PTGS2、CASP3、MAPK14因子的表达发挥止咳作用,为探究百部止咳的作用机制提供了理论参考。

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

国家自然科学基金面上项目（82174060）；黑龙江省自然科学基金资助项目（LH2021H074）；黑龙江省卫生健康委科研课题（20210202040011）