目的 基于网络药理学、分子对接以及体内外实验，探究地八角Astragalus bhotanensis治疗慢性咽炎的作用机制。方法 体外考察地八角提取物对金黄色葡萄球菌（Staphylococcus aureus，SA）、β-溶血性链球菌（β-hemolytic streptococcus，HS）、大肠埃希菌（Escherichia coli，EC）和肺炎双球菌（Diplococcus pneumoniae，DP）的抗菌活性。建立HS诱导的慢性咽炎大鼠模型，考察地八角提取物的药效作用。利用PubChem、GeneCards、STRING等数据库构建地八角提取物与慢性咽炎的相互作用网络，并借助Cytoscape软件筛选核心靶点，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。采用Autodock vina软件进行分子对接验证，并通过Western blotting验证预测结果。结果 地八角提取物对SA、HS、EC、DP均具有一定的抗菌活性。与模型组比较，地八角提取物可显著降低慢性咽炎大鼠血清中炎症因子水平（P＜0.05、0.01、0.001），并有效逆转咽部组织的病理损伤。网络药理学分析表明，地八角提取物可能通过调控丝裂原活化蛋白激酶（mitogen-activated protein kinase，MAPK）和磷脂酰肌醇3-激酶（phosphatidylinositol 3-kinase，PI3K）/蛋白激酶B（protein kinase B，Akt）等信号通路，作用于热休克蛋白90α家族B类成员1（heat shock protein 90 alpha family class B member 1，HSP90AB1）、磷脂酰肌醇-3-激酶催化亚基α（phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha，PIK3CA）、MAPK1、MAPK3、表皮生长因子受体（epidermal growth factor receptor，EGFR）和雌激素受体1（estrogen receptor 1，ESR1）等靶点，从而发挥治疗慢性咽炎的作用。分子对接结果表明，油酸、7β-羟基谷甾醇、astrabhotin A与MAPK蛋白具有良好的结合能力。Western blotting结果显示，地八角提取物可显著降低慢性咽炎大鼠咽部组织TLR4蛋白表达以及核因子-κB p65（nuclear factor-κB p65，NF-κB p65）和p38 MAPK的磷酸化水平（P＜0.05、0.01、0.001）。结论 地八角提取物通过调控TLR4/p38 MAPK/NF-κB p65信号通路缓解HS诱导的大鼠慢性咽炎

Objective To explore the mechanism of Astragalus bhotanensis (AB) against chronic pharyngitis based on network pharmacology, molecular docking, in vitro and in vivo experiments. Methods The antibacterial activity of AB extract against Staphylococcus aureus (SA), β-hemolytic streptococcus (HS), Escherichia coli (EC) and Diplococcus pneumoniae (DP) was assessed in vitro. A chronic pharyngitis rat model induced by HS was established and the pharmacological effects of AB extract was investigated. The interaction network between AB extract and chronic pharyngitis was constructed using databases such as PubChem, GeneCards, and STRING, and core targets were screened using Cytoscape software, followed by gene ontology (GO) function and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis. Molecular docking was performed using Autodock vina software, and the predicted results were verified by Western blotting. Results AB extract showed certain antibacterial activity against SA, HS, EC and DP. Compared with model group, AB extract significantly reduced the levels of inflammatory factors in serum of rats with chronic pharyngitis (P < 0.05, 0.01, 0.001), and effectively reversed the pathological damage of pharyngeal tissue. Network pharmacology analysis suggested that AB extract may act on targets such as heat shock protein 90 alpha family class B member 1 (HSP90AB1), phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA), MAPK1, MAPK3, epidermal growth factor receptor (EGFR) and estrogen receptor 1 (ESR1) by regulating signaling pathways such as mitogen activated protein kinase (MAPK) and phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt), thereby exerting therapeutic effects on chronic pharyngitis. The molecular docking results showed that oleic acid, 7β-hydroxysitosterol and astribhotin A had good binding ability with MAPK protein. Western blotting results showed that AB extract significantly reduced TLR4 protein expression and phosphorylation levels of nuclear factor-κB p65 (NF-κB p65) and p38 MAPK in pharyngeal tissue of chronic pharyngitis rats (P < 0.05, 0.01, 0.001). Conclusion AB extract alleviates HS-induced chronic pharyngitis in rats by regulating TLR4/p38 MAPK/NF-κB p65 signaling pathway.

R285.5

国家自然科学基金资助项目（82060707）