目的 探索大青龙汤抑制甲型流感病毒（H1N1/PR8）致小鼠病毒性肺炎的药效和机制。方法 通过UPLC-QE-HF-MS/MS检测大青龙汤中的主要化学成分。小鼠随机分为对照组、模型组、奥司他韦（27.5 mg/kg）组和大青龙汤高、中、低剂量（23.10、11.55、5.78 g/kg）组，建立甲型流感病毒（H1N1/PR8）感染致小鼠肺炎模型，连续给药4 d后，通过检测肺部病毒载量、肺组织炎症因子水平、外周血淋巴细胞比例、肺部病理结构与影像学，评价大青龙汤对病毒性肺炎的治疗作用，并通过蛋白质组学和Western blotting研究其作用机制。结果 鉴定出大青龙汤中77个主要化学成分。与模型组比较，大青龙汤显著降低感染小鼠肺组织病毒载量（P＜0.01），降低肺组织中肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）水平（P＜0.05、0.01），升高外周血中CD4⁺ T细胞比例（P＜0.05），明显改善肺部病变情况，并显著上调感染小鼠肺组织中活化T细胞核因子5（nuclear factor of activated T-cells 5，NFAT5）的表达（P＜0.05、0.01），下调酪氨酸蛋白激酶ZAP-70（tyrosine-protein kinase ZAP-70，ZAP70）的表达（P＜0.05）。结论 大青龙汤能够抑制感染小鼠肺组织中病毒增殖、降低TNF-α表达、提高CD4⁺ T淋巴细胞水平、改善肺部病变，其治疗病毒性肺炎的机制与调节NFAT5和ZAP70介导的T细胞免疫有关。

Objective To explore the efficacy and mechanism of Daqinglong Decoction (大青龙汤) in inhibiting viral pneumonia in mice caused by influenza A virus (H1N1/PR8). Methods The main chemical components in Daqinglong Decoction were detected by UPLC-QE-HF-MS/MS. Mice were randomly divided into control group, model group, oseltamivir (27.5 mg/kg) group, Daqinglong Decoction high-, medium-, and low-dose (23.10, 11.55, 5.78 g/kg) groups. The model of pneumonia in mice caused by influenza A virus (H1N1/PR8) infection was established, after continuously administration for 4 d, the therapeutic effect of Daqinglong Decoction on viral pneumonia was evaluated by detecting the viral load of lungs, the levels of inflammatory factors in lung tissues, the percentage of peripheral blood lymphocytes, and the pathological structure and imaging of lungs. The mechanism was studied through proteomics and Western blotting. Results A total of 77 major chemical constituents in Daqinglong Decoction were identified. Compared with model group, Daqinglong Decoction significantly reduced the viral load of lung tissues (P < 0.01), decreased the level of tumor necrosis factor-α (TNF-α) in lung tissues (P < 0.05, 0.01), increased the percentage of CD4⁺ T cells in the peripheral blood (P < 0.05), markedly improved lung lesions, and significantly up-regulated the expression of nuclear factor of activated T-cells 5 (NFAT5) and down-regulated the expression of tyrosine-protein kinase ZAP-70 (ZAP70) in lung tissues of infected mice (P < 0.05). Conclusion Daqinglong Decoction inhibited viral proliferation in lung tissues, decreased TNF-α expression, increased CD4⁺ T lymphocyte level and ameliorated lung lesions, and its mechanism of treating viral pneumonia was related to the modulation of NFAT5 and ZAP70 mediated T cell immunity.

R285.5

国家自然科学基金面上项目（82151210）；北京市自然科学基金项目（L222130）；中国中医科学院科技创新工程项目（CI2021A04607）