目的 采用網(wǎng)絡(luò)藥理學(xué)與分子對(duì)接技術(shù)探討熱毒寧注射液抗嚴(yán)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(jī)制與活性成分。方法 利用中藥系統(tǒng)藥理學(xué)數(shù)據(jù)庫(kù)和分析平臺(tái)（TCMSP）數(shù)據(jù)庫(kù)檢索熱毒寧注射液中青蒿、金銀花、梔子的主要化學(xué)成分及其作用靶點(diǎn)。運(yùn)用UniProt數(shù)據(jù)庫(kù)查詢(xún)相關(guān)靶點(diǎn)對(duì)應(yīng)的基因，通過(guò)Cytoscape 3.8.2構(gòu)建中藥材-化合物-靶點(diǎn)（基因）網(wǎng)絡(luò)；在GeneCards數(shù)據(jù)庫(kù)中搜集“COVID-19”“SARS”和“MERS”相關(guān)靶點(diǎn)，通過(guò)Venny 2.1.0數(shù)據(jù)庫(kù)映射篩選出3種冠狀病毒感染疾病的共有靶點(diǎn)；將SARS、MERS和COVID-19的共有靶點(diǎn)與熱毒寧注射液化合物靶點(diǎn)進(jìn)行交集篩選出共同靶點(diǎn)作為研究靶點(diǎn)；將共同靶點(diǎn)導(dǎo)入STRING數(shù)據(jù)庫(kù)獲取數(shù)據(jù)后，使用Cytoscape 3.8.2軟件構(gòu)建蛋白質(zhì)-蛋白質(zhì)相互作用（PPI）的網(wǎng)絡(luò)圖；利用R語(yǔ)言進(jìn)行基因本體論（GO）生物學(xué)功能富集分析及京都基因和基因組百科全書(shū)（KEGG）信號(hào)通路富集分析，繪制柱狀圖及氣泡圖進(jìn)行可視化分析，并構(gòu)建成分-靶點(diǎn)-通路網(wǎng)絡(luò)圖；選取成分-靶點(diǎn)-通路網(wǎng)絡(luò)中關(guān)鍵化合物與重要靶點(diǎn)蛋白及新型冠狀病毒（SARS-CoV-2）3CL水解酶、血管緊張素轉(zhuǎn)化酶II（ACE2）進(jìn)行分子對(duì)接。結(jié)果 熱毒寧注射液篩選得到31個(gè)活性化合物中，207個(gè)相應(yīng)作用靶點(diǎn)；SARS相關(guān)靶點(diǎn)2 453個(gè)，MERS相關(guān)靶點(diǎn)805個(gè)，COVID-19相關(guān)靶點(diǎn)2 571個(gè)，3種疾病共有靶點(diǎn)786個(gè)，與熱毒寧注射液的共同靶點(diǎn)11個(gè)，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(jìn)程（BP） 995個(gè)，分子功能（MF） 71種，細(xì)胞組分（CC）31種。KEGG通路富集分析篩選得到99條信號(hào)通路（P＜0.05），主要涉及前列腺癌、流體剪切應(yīng)力和動(dòng)脈粥樣硬化、肝細(xì)胞癌、癌癥中的蛋白多糖、脂質(zhì)與動(dòng)脈粥樣硬化、人類(lèi)T細(xì)胞白血病病毒1型感染、MAPK信號(hào)通路等。分子對(duì)接結(jié)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(lèi)化合物與關(guān)鍵靶點(diǎn)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(jié)合能較推薦化學(xué)藥小。結(jié)論 熱毒寧注射液對(duì)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(diǎn)，調(diào)節(jié)多種信號(hào)通路發(fā)揮抑制炎癥風(fēng)暴、調(diào)節(jié)免疫功能、抗病毒等作用有關(guān)。

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(xué)科技攻關(guān)計(jì)劃項(xiàng)目(LHGJ20200402)