目的 通过网络药理学结合体内实验验证，探讨茅岩莓Ampelopsis grossedentata提取物治疗感染后咳嗽（post-infection cough，PIC）的作用及机制。方法 通过HERB数据库筛选茅岩莓活性成分，结合GeneCards平台获取PIC相关靶点，得到交集靶点并构建“药物-活性成分-靶点”网络及蛋白质-蛋白质相互作用（protein-protein interaction，PPI）网络，进行基因本体（gene ontology，GO）功能及京都基因与基因组百科全书（Kyoto encyclopedia of genes and genomes，KEGG）通路富集分析，同时对核心成分与核心靶点进行分子对接验证。构建PIC豚鼠模型，设置对照组、模型组及茅岩莓提取物低、中、高剂量（64、128、256 mg/kg）组和苏黄止咳胶囊（314 mg/kg）组，给药后检测咳嗽敏感性、肺泡灌洗液中炎性细胞数量及白细胞介素-4（interleukin-4，IL-4）、P物质（substance P，SP）、γ干扰素（interferon-γ，IFN-γ）、肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）水平，采用苏木素-伊红（hematoxylin-eosin，HE）染色观察气管、支气管及肺组织病理变化，采用Western blotting检测肺组织中核心靶点蛋白的表达。结果 网络药理学筛选出茅岩莓核心活性成分包括大黄素甲醚、大黄素、槲皮素等，核心靶点有B细胞淋巴瘤-2（B-cell lymphoma-2，Bcl-2）、表皮生长因子受体（epidermal growth factor receptor，EGFR）、信号转导与转录激活因子3（signal transducer and activator of transcription 3，STAT3）、雌激素受体1（estrogen receptor 1，ESR1）、肉瘤原癌基因激酶（sarcoma proto-oncogene kinase，SRC）等，富集涉及EGFR、磷脂酰肌醇3-激酶（phosphatidylinositol 3-kinase，PI3K）-蛋白激酶B（protein kinase B，Akt）等信号通路；分子对接显示核心成分与核心靶点结合能较低，亲和力良好。动物实验结果显示，与模型组比较，茅岩莓提取物可显著延长豚鼠咳嗽潜伏期并减少咳嗽次数（P＜0.05、0.01），降低肺泡灌洗液中炎性细胞数量及SP、IL-4、TNF-α、INF-γ水平（P＜0.05、0.01），改善气管、支气管及肺组织病理损伤，上调肺组织Bcl-2蛋白表达（P＜0.01），下调EGFR、p-STAT3蛋白表达（P＜0.01）。结论 茅岩莓提取物对PIC具有显著的治疗效果，其作用机制可能为通过大黄素甲醚、大黄素、槲皮素等核心成分作用于Bcl-2、EGFR、STAT3等靶点，调控相关信号通路，减轻气道黏膜损伤及炎症反应，从而发挥治疗作用。

Objective To investigate the effect and mechanism of Ampelopsis grossedentata extract in treating post-infection cough (PIC) through network pharmacology combined with in vivo experiments. Methods Active components from A. grossedentata were screened through HERB database, PIC related targets were obtained from GeneCards platform, intersecting targets were obtained, “drug-active ingredient-target” network and protein-protein interaction (PPI) network were constructed. Gene ontology (GO) function and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis were performed, and molecular docking validation was performed between core components and core targets. PIC guinea pig model was constructed, control group, model group, A. grossedentata extract low-, medium-, high-dose (64, 128, 256 mg/kg) groups and Suhuang Zhike Capsule (苏黄止咳胶囊, 314 mg/kg) group were set up. After administration, cough sensitivity, inflammatory cell numbers and levels of interleukin-4 (IL-4), substance P (SP), interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α) in bronchoalveolar lavage fluid were detected. Hematoxylin-eosin (HE) staining was used to observe pathological changes in tracheal, bronchial and lung tissues. Western blotting was used to detect the expressions of core target proteins in lung tissue. Results The core active ingredients of A. grossedentata screened by network pharmacology included physcion, emodin, quercetin, etc. The core targets included B-cell lymphoma-2 (Bcl-2), epidermal growth factor receptor (EGFR), signal transducer and activator of transcription 3 (STAT3), estrogen receptor 1 (ESR1), sarcoma proto oncogene kinase (SRC), etc. The enrichment involved EGFR, phosphatidylinositol 3-kinase (PI3K)-protein kinase B (Akt) and other signaling pathways. Molecular docking showed that the binding energy between core components and core targets was low, with good affinity. The animal experiment results showed that compared with model group, A. grossedentata extract could significantly prolong the cough latency and reduce the number of coughs in guinea pigs (P < 0.05, 0.01), reduce the number of inflammatory cells and levels of SP, IL-4, TNF-α, INF-γ in bronchoalveolar lavage fluid (P < 0.05, 0.01), improve the pathological damage of trachea, bronchus and lung tissue, up-regulate the expression of Bcl-2 protein in lung tissue (P < 0.01), down-regulate the expressions of EGFR and p-STAT3 protein (P < 0.01). Conclusion A. grossedentata extract has significant therapeutic effects on PIC, and its mechanism may be through the action of core components such as physcion, emodin and quercetin on targets such as Bcl-2, EGFR, STAT3, regulating related signaling pathways, reducing airway mucosal damage and inflammatory reactions, and thus exerting therapeutic effects.

R285.5

湖南省科技创新创业团队（2021）；湖南省科技领军人才项目（2024）