目的 采用網絡藥理學與分子對接技術探討熱毒寧注射液抗嚴重急性呼吸綜合征（SARS）、中東呼吸綜合征（MERS）和新型冠狀病毒肺炎（COVID-19）的潛在共性作用機制與活性成分。方法 利用中藥系統(tǒng)藥理學數據庫和分析平臺（TCMSP）數據庫檢索熱毒寧注射液中青蒿、金銀花、梔子的主要化學成分及其作用靶點。運用UniProt數據庫查詢相關靶點對應的基因，通過Cytoscape 3.8.2構建中藥材-化合物-靶點（基因）網絡；在GeneCards數據庫中搜集“COVID-19”“SARS”和“MERS”相關靶點，通過Venny 2.1.0數據庫映射篩選出3種冠狀病毒感染疾病的共有靶點；將SARS、MERS和COVID-19的共有靶點與熱毒寧注射液化合物靶點進行交集篩選出共同靶點作為研究靶點；將共同靶點導入STRING數據庫獲取數據后，使用Cytoscape 3.8.2軟件構建蛋白質-蛋白質相互作用（PPI）的網絡圖；利用R語言進行基因本體論（GO）生物學功能富集分析及京都基因和基因組百科全書（KEGG）信號通路富集分析，繪制柱狀圖及氣泡圖進行可視化分析，并構建成分-靶點-通路網絡圖；選取成分-靶點-通路網絡中關鍵化合物與重要靶點蛋白及新型冠狀病毒（SARS-CoV-2）3CL水解酶、血管緊張素轉化酶II（ACE2）進行分子對接。結果 熱毒寧注射液篩選得到31個活性化合物中，207個相應作用靶點；SARS相關靶點2 453個，MERS相關靶點805個，COVID-19相關靶點2 571個，3種疾病共有靶點786個，與熱毒寧注射液的共同靶點11個，分別為HSPA5、CRP、MAPK1、HMOX1、TGFB1、HSP90AA1、TP53、DPP4、CXCL10、PLAT、PRKACA。GO功能富集分析得到生物進程（BP） 995個，分子功能（MF） 71種，細胞組分（CC）31種。KEGG通路富集分析篩選得到99條信號通路（P＜0.05），主要涉及前列腺癌、流體剪切應力和動脈粥樣硬化、肝細胞癌、癌癥中的蛋白多糖、脂質與動脈粥樣硬化、人類T細胞白血病病毒1型感染、MAPK信號通路等。分子對接結果顯示熱毒寧注射液中槲皮素、木犀草素、山柰酚3種核心活性黃酮類化合物與關鍵靶點MAPK1、PRKACA、HSP90AA1具有很好的親和力，并且3種活性化合物與SARS-CoV-2 3CL水解酶及ACE2結合能較推薦化學藥小。結論 熱毒寧注射液對SARS、MERS和COVID-19 3種疾病具有潛在共性作用，該作用可能與活性化合物槲皮素、木犀草素、山柰酚等作用于MAPK1、PRKACA、HSP90AA1等靶點，調節(jié)多種信號通路發(fā)揮抑制炎癥風暴、調節(jié)免疫功能、抗病毒等作用有關。

Objective To explore the potential common mechanism and active ingredients of Reduning Injection against SARS, MERS and COVID-19 through network pharmacology and molecular docking technology. Methods The TCMSP database was used to retrieve the chemical components and targets of Artemisiae Annuae Herba, Lonicerae Japonicae Flos and Gardeniae Fructus in Reduning Injection. The gene corresponding to the target was searched by UniProt database, and Cytoscape 3.8.2 was used to build a medicinal material-compound-target (gene) network. Three coronavirus-related targets were collected in the Gene Cards database with the key words of "SARS""MERS" and "COVID-19", and common target of three coronavirus infection diseases were screened out through Venny 2.1.0 database. The common targets of SARS, MERS and COVID-19 were intersected with the targets of Reduning Injection, and the common targets were selected as research targets. Protein-protein interaction (PPI) network map were constructed by Cytoscape3.8.2 software after importing the common targets into the STRING database to obtain data. R language was used to carry out GO biological function enrichment analysis and KEGG signaling pathway enrichment analysis, histograms and bubble charts were drew, and component-target-pathway network diagrams was constructed. The key compounds in the component-target-pathway network were selected for molecular docking with important target proteins, novel coronavirus (SARS-CoV-2) 3CL hydrolase, and angiotensin-converting enzyme II (ACE2). Results 31 active compounds and 207 corresponding targets were obtained from Reduning Injection. 2 453 SARS-related targets, 805 MERS-related targets, 2 571 COVID- 19-related targets, and 786 targets for the three diseases. 11 common targets with Reduning Injection: HSPA5, CRP, MAPK1, HMOX1, TGFB1, HSP90AA1, TP53, DPP4, CXCL10, PLAT, PRKACA. GO function enrichment analysis revealed 995 biological processes (BP), 71 molecular functions (MF), and 31 cellular components (CC). KEGG pathway enrichment analysis screened 99 signal pathways (P<0.05), mainly related to prostate cancer, fluid shear stress and atherosclerosis, hepatocellular carcinoma, proteoglycans in cancer, lipid and atherosclerosis, human T-cell leukemia virus 1 infection, MAPK signaling pathway, etc. The molecular docking results showed that the three core active flavonoids of quercetin, luteolin, and kaempferol in Reduning Injection had good affinity with key targets MAPK1, PRKACA, and HSP90AA1, and the combination of the three active compounds with SARS-CoV-2 3CL hydrolase and ACE2 was less than the recommended chemical drugs. Conclusion Reduning Injection has potential common effects on the three diseases of SARS, MERS and COVID-19. This effect may be related to those active compounds such as quercetin, luteolin, and kaempferol acting on targets such as MAPK1, PRKACA, HSP90AA1 to regulate multiple signal pathways and exert anti-virus, suppression of inflammatory storm, and regulation of immune function.

R285.5

河南省醫(yī)學科技攻關計劃項目(LHGJ20200402)