目的 明确病炎清的抗流感病毒活性，基于网络药理学和实验验证探究病炎清治疗流感的潜在机制。 方法 检测病炎清对流感模型小鼠死亡率、肺指数、肺病毒量的影响。检索 TCMSP、Swiss Target Prediction 等 数据库及文献，收集病炎清处方成分及靶点；通过 Cytoscape 软件构建病炎清抗流感“成分-共有靶点”网络； 通过 STRING 数据库和 Cytoscape 软件构建蛋白互作（Protein-Protein Interaction，PPI）网络，筛选核心靶点；对 共有靶点进行基因本体（Gene Ontology，GO）和京都基因组百科全书（Kyoto Encyclopedia of Genes and Genomes， KEGG）富集分析。在体内水平探究病炎清减轻流感所致氧化损伤的作用，以及对抗氧化和促氧化酶表达、P65 蛋白磷酸化、核因子 E2 相关因子 2（Nuclear Factor Erythroid-Derived 2-Related Factor 2，Nrf2）核易位的影响。 结果 病炎清可明显降低流感模型小鼠死亡率，降低肺指数和肺病毒量，减轻肺损伤。检索得到病炎清 193 个 活性成分，对应 338 个靶点，其中与流感相关的靶点 180 个，IκBα 激酶 α（CHUK）、IκBα 激酶 γ（IKBKG）、 NF-κB p65（RELA）、肿瘤坏死因子（TNF）、白介素 6（IL6）等 9 个靶点为潜在核心靶点。GO 分析提示病炎清具 有抗菌、抗氧化应激等多种生物功能。KEGG 分析表明病炎清涉及多条病毒及免疫通路，如疱疹病毒、甲型流 感病毒、肿瘤坏死因子和 Toll 样受体途径。动物实验表明，相较于磷酸奥司他韦，病炎清高剂量组可更明显 地降低流感模型小鼠肺丙二醛（Malondialdehyde，MDA）含量（P＜0.01）、提升肺总超氧化物歧化酶（Superoxide Dismutase，SOD）活性（P＜0.001）。相较于磷酸奥司他韦，病炎清&磷酸奥司他韦联合给药组肺部具有更低的 NADPH 氧化酶 2（NADPH Oxidase Type 2，NOX2）、4（NOX4）水平（P＜0.001）和更高的过氧化氢酶（Catalase， CAT）、谷胱甘肽过氧化物酶 4（Glutathione Peroxidase 4，GPX4）水平（P＜0.001）。另外，联合给药组显示出更 强的 Nrf2 核易位（P＜0.05）。但联合给药组 P65 蛋白磷酸化水平与磷酸奥司他韦组比较，差异无统计学意义 （P＞0.05），但均低于模型组（P＜0.05）。结论 病炎清具有明显的抗流感病毒活性，可发挥抑制促氧化酶合成 和促进抗氧化系统的双重作用，减轻流感引发的氧化应激损伤。

Objective This study aimed to clarify the anti-influenza virus activity of Bingyanqing （BYQ），as well as to explore the mechanism of BYQ in treating influenza through network pharmacology and experimental verification. Methods The impact of BYQ on mortality，lung index，and viral load in an influenza mouse model was detected. We collected the ingredients and targets of BYQ formula by searching databases including TCMSP，Swiss Target Prediction and consulting the literature. The“ingredients -common target”network for anti-influenza effect of BYQ was constructed using Cytoscape software. The protein-protein interaction （PPI） network was constructed using the STRING database and Cytoscape software，and the core targets were screened. Gene Ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） enrichment analyses were performed for common targets. The effect of BYQ on reducing influenza-induced oxidative damage， the expression of antioxidant and pro-oxidant enzyme， P65 phosphorylation and nuclear factor E2-related factor 2 （Nrf2） nuclear translocation in vivo were investigated. Results BYQ significantly reduced mortality，lung index，pulmonary viral load and lung injury in a mouse model of influenza. We obtained one hundred and ninety-three of BYQ active compounds， which corresponded to three hundred and thirty-eight targets. There are 180 influenza-related targets among them. Nine targets，including IκBα kinase α （CHUK），IκBα kinase γ （IKBKG），NF- κB p65 （RELA），tumor necrosis factor （TNF） and interleukin 6 （IL6），were identified as potential core targets. GO analysis indicated that BYQ is involved in several biological functions， including antibacterial and antioxidant stress responses. KEGG analysis revealed the involvement of several viral and immune-related pathways for BYQ in treating influenza， including herpes simplex virus，influenza A virus，TNFα and toll-like receptor pathways. In vivo studies showed that highdose BYQ significantly reduced pulmonary malondialdehyde （MDA） levels （P＜0.01） and increased total superoxide dismutase （SOD）activity （P＜0.001）in a mouse mode of influenza compared to oseltamivir phosphate. The treatment group with the combination of BYQ & oseltamivir phosphate had lower levels of NADPH oxidase 2（NOX2）and 4（NOX4）（P＜0.001），and higher levels of catalase （CAT） and glutathione peroxidase 4（GPX4） （P＜0.001） in the lungs than oseltamivir phosphate group. The combined treatment group showed more significant Nrf2 nuclear translocation （P＜0.05） than the oseltamivir phosphate group. However， there was no significant difference in P65 phosphorylation levels between the combination treatment group and the oseltamivir phosphate group （P＜0.05）， but P65 phosphorylation levels in both groups were lower than in the model group （P＜0.05）. Conclusion BYQ exhibits significant anti-influenza virus activity，manifests a dual effect by inhibiting the synthesis of pro-oxidant enzymes and promoting the antioxidant system，thereby alleviates the oxidative stress damage caused by influenza.

R285.5

国家自然科学基金青年基金（81904103）；深圳市科技计划基础研究面上项目（JCYJ20190812155412785，JCYJ20210324111204013）； 山东省自然基金青年基金（ZR2020QH328）。