目的 采用网络药理学与分子对接技术探讨热毒宁注射液抗严重急性呼吸综合征（SARS）、中东呼吸综合征（MERS）和新型冠状病毒肺炎（COVID-19）的潜在共性作用机制与活性成分。方法 利用中药系统药理学数据库和分析平台（TCMSP）数据库检索热毒宁注射液中青蒿、金银花、栀子的主要化学成分及其作用靶点。运用UniProt数据库查询相关靶点对应的基因，通过Cytoscape 3.8.2构建中药材-化合物-靶点（基因）网络；在GeneCards数据库中搜集“COVID-19”“SARS”和“MERS”相关靶点，通过Venny 2.1.0数据库映射筛选出3种冠状病毒感染疾病的共有靶点；将SARS、MERS和COVID-19的共有靶点与热毒宁注射液化合物靶点进行交集筛选出共同靶点作为研究靶点；将共同靶点导入STRING数据库获取数据后，使用Cytoscape 3.8.2软件构建蛋白质-蛋白质相互作用（PPI）的网络图；利用R语言进行基因本体论（GO）生物学功能富集分析及京都基因和基因组百科全书（KEGG）信号通路富集分析，绘制柱状图及气泡图进行可视化分析，并构建成分-靶点-通路网络图；选取成分-靶点-通路网络中关键化合物与重要靶点蛋白及新型冠状病毒（SARS-CoV-2）3CL水解酶、血管紧张素转化酶II（ACE2）进行分子对接。结果 热毒宁注射液筛选得到31个活性化合物中，207个相应作用靶点；SARS相关靶点2 453个，MERS相关靶点805个，COVID-19相关靶点2 571个，3种疾病共有靶点786个，与热毒宁注射液的共同靶点11个，分别为HSPA5、CRP、MAPK1、HMOX1、TGFB1、HSP90AA1、TP53、DPP4、CXCL10、PLAT、PRKACA。GO功能富集分析得到生物进程（BP） 995个，分子功能（MF） 71种，细胞组分（CC）31种。KEGG通路富集分析筛选得到99条信号通路（P＜0.05），主要涉及前列腺癌、流体剪切应力和动脉粥样硬化、肝细胞癌、癌症中的蛋白多糖、脂质与动脉粥样硬化、人类T细胞白血病病毒1型感染、MAPK信号通路等。分子对接结果显示热毒宁注射液中槲皮素、木犀草素、山柰酚3种核心活性黄酮类化合物与关键靶点MAPK1、PRKACA、HSP90AA1具有很好的亲和力，并且3种活性化合物与SARS-CoV-2 3CL水解酶及ACE2结合能较推荐化学药小。结论 热毒宁注射液对SARS、MERS和COVID-19 3种疾病具有潜在共性作用，该作用可能与活性化合物槲皮素、木犀草素、山柰酚等作用于MAPK1、PRKACA、HSP90AA1等靶点，调节多种信号通路发挥抑制炎症风暴、调节免疫功能、抗病毒等作用有关。

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

河南省医学科技攻关计划项目(LHGJ20200402)