目的 基于网络药理学和实验验证探究青龙白虎汤改善急性咽炎的作用机制。方法 通过网络药理学预测青龙白虎汤改善急性咽炎的生物调控过程和信号通路，构建蛋白质-蛋白质相互作用（protein-protein interaction，PPI）网络和“成分-通路”网络。建立氨水诱导的急性咽炎大鼠模型，给予青龙白虎汤干预后，测定大鼠体质量、咽喉组织指数和肺指数；采用苏木素-伊红（HE）染色观察咽喉组织病理变化；采用试剂盒检测血清中肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）、白细胞介素-1β（interleukin-1β，IL-1β）、IL-6、前列腺素E₂（prostaglandin E₂，PGE₂）水平以及血清和咽喉组织中丙二醛（malondialdehyde，MDA）水平和超氧化物歧化酶（superoxide dismutase，SOD）、谷胱甘肽过氧化物酶（glutathione peroxidase，GSH-Px）活性；采用Western blotting检测咽喉组织中Toll样受体4（Toll-like receptor 4，TLR4）/核因子-κB（nuclear factor-κB，NF-κB）和核因子E2相关因子2（nuclear factor erythroid 2-related factor 2，Nrf2）/血红素氧合酶-1（heme oxygenase-1，HO-1）信号通路相关蛋白表达。结果 共获得青龙白虎汤中26个活性成分，86个活性成分与疾病的交集靶点，其中核心靶点34个。网络药理学分析结果显示，青龙白虎汤可以调节TNF-α、IL-1β、IL-6等核心靶点，调节抗氧化等生物过程，调控NF-κB信号通路，从而治疗急性咽炎。体内实验结果显示，青龙白虎汤能够改善急性咽炎大鼠咽部组织损伤，显著抑制血清中炎症因子水平（P＜0.05、0.01、0.001），调控血清和咽喉组织中的氧化应激水平（P＜0.05、0.01、0.001），显著下调TLR4/NF-κB信号通路相关蛋白表达（P＜0.05、0.01、0.001），显著上调Nrf2/HO-1信号通路相关蛋白表达（P＜0.05、0.001）。结论 青龙白虎汤能够通过抑制TLR4/NF-κB信号通路并激活Nrf2/HO-1信号通路，抑制炎症反应，调控氧化应激，从而改善急性咽炎。

Objective To explore the mechanism of Qinglong Baihu Decoction (青龙白虎汤, QBD) on improving acute pharyngitis based on network pharmacology and experimental verification. Methods The biological regulatory processes and signaling pathways of QBD on improving acute pharyngitis were predicted by network pharmacology, protein-protein interaction (PPI) network and “component-pathway” network were constructed. A rat model of acute pharyngitis induced by ammonium hydroxide was established, and after intervention with QBD, body weight, pharyngeal tissue index and lung index of rats were measured; Hematoxylin-eosin (HE) staining was used to observe pathological changes in pharyngeal tissue; Reagent kit was used to detect the levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), IL-6, prostaglandin E₂ (PGE₂) in serum, as well as the level of malondialdehyde (MDA) and activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in serum and pharyngeal tissue; Western blotting was used to detect the expressions of Toll-like receptor 4 (TLR4)/nuclear factor-κB (NF-κB) and nuclear factor erythroid 2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) signaling pathway related proteins in pharyngeal tissue. Results A total of 26 active ingredients were obtained from QBD, and 86 interaction targets between active ingredients and diseases, including 34 core targets. The results of network pharmacology analysis showed that QBD could regulate core targets such as TNF-α, IL-1β and IL-6, regulate biological processes such as antioxidant activity, and regulate NF-κB signaling pathway, thereby treating acute pharyngitis. The in vivo experimental results showed that QBD could improve the damage of pharyngeal tissue in rats with acute pharyngitis, significantly inhibit the levels of inflammatory factors in serum (P < 0.05, 0.01, 0.001), regulate the levels of oxidative stress in serum and pharyngeal tissue (P < 0.05, 0.01, 0.001), significantly down-regulate the expressions of TLR4/NF-κB signaling pathway related proteins (P < 0.05, 0.01, 0.001), and significantly up-regulate the expressions of Nrf2/HO-1 signaling pathway related proteins (P < 0.05, 0.001). Conclusion QBD can improve acute pharyngitis by inhibiting TLR4/NF-κB signaling pathway and activating Nrf2/HO-1 signaling pathway, suppressing inflammatory responses and regulating oxidative stress.

R285.5

四川省中医药管理局中医药开发专项（2018KF001）；成都市科技局国际科技合作项目（2022-GH02-00033-HZ）