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.

Astragalus bhotanensis Baker  /  chronic pharyngitis  /  antibiosis  /  anti-inflammatory  /  TLR4/p38 MAPK/NF-κB signaling pathway  /  liquiritigenin  /  isoliquiritigenin  /  carotenoids