Investigating the Potential Targets and Mechanisms of Action of Chinese Medicine for the Treatment of EIV Based on Network Pharmacology and Molecular Docking as Well as <i>In Vitro</i>

Background Equine influenza virus (EIV) can cause acute infections and outbreaks of epidemics in horses and donkeys. It is one of the most economically impactful pathogens in equine respiratory diseases globally, resulting in substantial financial losses within the farming industry. Utilizing targeted anti-viral drugs is an effective strategy. Purpose The present study analyzes the potential of herbal medicines for the treatment of equine influenza (EI) based on network pharmacology, molecular docking techniques, and in vitro anti-viral studies. Materials and Methods The construction of a “traditional Chinese medicine (TCM) component–target–disease” network was performed using Cytoscape 3.9.0. The protein–protein interaction (PPI) network is performed through the STRING system. Bioconductor software was employed to conduct gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) functional enrichment analyses of biological processes (BPs). Molecular docking techniques revealed the degree of binding of core components to key target genes. Characterization of the anti-EI effect of TCM by cytotoxicity and in vitro studies. Results Consequently, five core TCMs were screened, which had 79 core targets related to EI. PPI network analysis highlighted 10 significant targets. Molecular docking results analysis revealed binding interactions between the main core component, kaempferol, and the targets prostaglandin-endoperoxide synthase 2 (PTGS2), matrix metalloproteinase-9 (MMP9), and estimated glomerular filtration rate (EGFR), with binding energies of −9.1, −8.3, and −8.0 (kcal/mol), respectively. In vitro studies have demonstrated that the inhibitory effect of kaempferol on EI is mainly in the initial phase. Conclusion Through network pharmacology, molecular docking, and in vitro experiments, kaempferol was demonstrated to combat EI through key targets of PTGS2, MMP9, EGFR, AKT, tumor necrosis factor (TNF), and IL-6. This study provides a basis for treating EI with herbal medicine and for later drug development. Future research should integrate network pharmacology with clinical applications, focusing on large-scale clinical trials to evaluate the efficacy and safety of TCM in influenza treatment, thereby enhancing its potential role in treating the disease.