目的 基于網(wǎng)絡(luò)藥理學(xué)與分子對接探討清肺達原顆粒治療新型冠狀病毒肺炎（COVID-19）的主要活性成分、靶標(biāo)及通路，以期闡述其作用機制。方法 通過TCMSP、ETCM、YATCM數(shù)據(jù)庫檢索清肺達原顆粒的中藥化學(xué)成分，以口服生物利用度（OB）≥30%和類藥性（DL）≥0.18為閾值篩選潛在的活性化合物。通過SIB、STITCH數(shù)據(jù)庫查詢活性化合物對應(yīng)的靶標(biāo)，利用STRING數(shù)據(jù)庫獲取蛋白-蛋白相互作用（PPI）網(wǎng)絡(luò)和網(wǎng)絡(luò)拓?fù)鋮?shù)，導(dǎo)入Cytoscape 3.6.0對網(wǎng)絡(luò)進行分析，篩選關(guān)鍵靶標(biāo)。通過DAVID進行基因本體（GO）功能富集分析、京都基因與基因組百科全書（KEGG）通路富集分析和組織富集（tissue enrichment）預(yù)測其作用機制。利用AutoDock將化合物與新型冠狀病毒（SARS-CoV-2）S蛋白受體結(jié)合結(jié)構(gòu)域與血管緊張素轉(zhuǎn)化酶II（ACE2）蛋白酶結(jié)構(gòu)域復(fù)合物（SARS-CoV-2-S-RBD-ACE2）進行分子對接。結(jié)果 篩選得到清肺達原顆粒活性化合物251個，對應(yīng)靶點共1 037個；篩選出關(guān)鍵靶標(biāo)107個，對應(yīng)化合物185個，關(guān)鍵靶點涉及ESR1、AR、EGFR、KDR、MMP2等，與ACE2共表達基因52個。GO功能富集分析結(jié)果顯示清肺達原顆粒主要對細(xì)胞表面信號轉(zhuǎn)導(dǎo)、分子功能、磷酸化和轉(zhuǎn)錄等生物學(xué)過程起調(diào)節(jié)作用，KEGG通路富集主要涉及趨化因子信號通路、T細(xì)胞受體信號通路、B細(xì)胞受體信號通路、自然殺傷細(xì)胞介導(dǎo)的細(xì)胞毒性和Toll樣受體信號通路等。組織富集顯示關(guān)鍵基因表達部位主要分布在肺及上皮細(xì)胞，涉及多種免疫細(xì)胞，如T細(xì)胞、B細(xì)胞、淋巴細(xì)胞等。分子對接結(jié)果顯示與SARS-CoV-2-S-RBD-ACE2復(fù)合物親和力較好的化合物主要來源于柴胡、黨參、知母、甘草，其中柴胡皂苷、甘草酸、知母皂苷與SARS-CoV-2- S-RBD-ACE2復(fù)合物結(jié)合力較強，可能為抗SARS-CoV-2的潛在活性成分。結(jié)論 清肺達原顆粒具有多成分、多靶點、多途徑的整體調(diào)控特點。其中柴胡皂苷、甘草酸、知母皂苷可能為抗SARS-CoV-2的潛在活性成分，治療COVID-19的作用機制可能與調(diào)控ACE2共表達的基因、調(diào)節(jié)炎癥和免疫相關(guān)的信號通路、影響SARS-CoV-2-S-RBD-ACE2復(fù)合物的穩(wěn)定有關(guān)。

Objective To explore the main active components, key targets and signaling pathways of Qingfei Dayuan Granules in treating of COVID-19 based on network pharmacology and molecular docking. Methods TCMSP, ETCM and YATCM databases were used to search the chemical constituents of Qingfei Dayuan Granules, and the threshold values of OB ≥ 30% and DL ≥ 0.18 were used to screen the potential active compounds. SIB and STITCH databases were used to query the targets corresponding to the active compounds, and the PPI network and network topology parameters were obtained by using STRING database. Cytoscape 3.6.0 was used to screen the hub targets. The key targets were analyzed by Gene Ontology (GO), the Kyoto Encyclopedia of genes and genomes (KEGG) pathway enrichment and tissue enrichment using DAVID 6.8 software. The molecular docking was performed by AutoDock Tools 1.5.6 software. Results A total of 251 active compounds and 1 037 targets were obtained, 107 key targets and 185 corresponding compounds were screened. The key targets involved ESR1, AR, EGFR, KDR, MMP2, and 52 genes were coexpressed with ACE2. The results of GO function enrichment analysis showed that Qingfei Dayuan Granules mainly regulated the biological processes of cell surface signaling transduction, molecular function, phosphorylation and transcription. KEGG pathway enrichment mainly involved chemokine signaling pathway, T cell receptor signaling pathway, B cell receptor signaling pathway, natural killer cell mediated cytotoxicity and Toll like receptor signaling pathway. The results of tissue enrichment showed that the key gene expression sites were mainly in lung and epithelial cells, involving a variety of immune cells, such as T cells, B cells, lymphocytes, etc. Molecular docking showed that the compounds with good binding power to SARS-CoV-2-RBD-ACE2 complex in Qingfei Dayuan granules were mainly come from Bupleuri Radix, Codonopsis Radix, Anemarrhenae Rhizoma, and Glycyrrhizae Radix et Rhizoma. Saikosaponin, glycyrrhizic acid, anemarsaponin had good binding power with SARS-CoV-2-S-RBD-ACE2, which may be potential active components against SARS-CoV-2. Conclusion Qingfei Dayuan Granules has the characteristics of multi-components, multi-targets and multi-pathway regulation. Saikosaponin, glycyrrhizic acid, and anemarsaponin may be the potential active components against SARS-CoV-2. The mechanisms of its treatment against COVID-19 may be related to the regulation of the co-expressed genes with ACE2, inhibition of inflammation and immune related signaling pathways, and the destruction of the complex structure of SARS-CoV-2-S-RBD-ACE2.

R285.5

湖北省科技廳新型肺炎應(yīng)急科技攻關(guān)項目：基于肺炎1號為主的中醫(yī)藥治療新型冠狀病毒肺炎的臨床研究（2020FCA007）