Objective To explore the mechanism of Bukan Yili Decoction for treating gastroesophageal reflux disease (GERD) based on network pharmacology, and to verify it through molecular docking technology and animal experiments. Methods (1) The effective active components of Bukan Yili Decoction were obtained by searching the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP), the SymMap database, and literature. The effect targets of these active components were searched in the TCMSP and the SwissTargetPrediction database. GERD⁃related targets were obtained from the GeneCards® database, OMIM® database, Therapeutic Target Database, and Comparative Toxicogenomics Database. Venny 2.1 database was used to obtain the intersection targets between the effect targets of active components and disease⁃related targets. Cytoscape software was adopted to construct a drug⁃component⁃effect target network, which was analyzed with the NetworkAnalyzer tool, and the top 5 components ranked by degree value were selected as key active components. For the intersection targets, protein⁃protein interaction network analysis (to obtain core targets), Gene Ontology (GO) functional enrichment analysis, and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were performed. Molecular docking was performed to verify the binding of key active components with core target proteins. (2) Twenty⁃nine rats were randomly divided into modeling group or sham operation group. Rats in the modeling group underwent duodenal anastomosis to establish a GERD model, while rats in the sham operation group received only the same incision, with the muscle and skin sutured immediately after exposing the esophagogastric junction, and were fed normally after surgery. Rats with successful modeling were randomly divided into model group, or high⁃ , medium⁃, and low⁃dose Bukan Yili Decoction groups. Rats in the high⁃, medium⁃, and low⁃dose Bukan Yili Decoction groups received intragastric administration of Bukan Yili Decoction at 35.4 g/(kg·d), 17.7 g/(kg·d), and 8.85 g/(kg·d), respectively, while those in the sham operation group and the model group received intragastric administration of an equal volume of normal saline, twice daily for 2 consecutive weeks. The body weight, feces, food intake, activity, response sensitivity, and mental state of rats in each group were observed and recorded in detail daily. After the end of intervention, tissues from the middle and lower esophagus at the esophagogastric junction were collected from rats in each group. The pathomorphological changes were observed by HE staining, and the protein expressions of Jun proto⁃oncogene (JUN) and RELA proto⁃oncogene (RELA) in the tissues from the middle and lower esophagus of rats in each group were compared. Results (1) There were 189 effective active components of Bukan Yili Decoction, 689 effect targets of active components, and 752 GERD⁃related targets, with 125 intersection targets between them. Quercetin, β⁃sitosterol, kaempferol, luteolin, and naringenin were the key active components of Bukan Yili Decoction. Signal transducer and activator of transcription 3 (STAT3), JUN, RELA, interleukin 6 (IL⁃6), tumor protein p53 (TP53), and tumor necrosis factor (TNF) were the core targets of Bukan Yili Decoction in treating GERD. The intersection targets were mainly involved in biological processes such as response to xenobiotic stimulus and cellular response to chemical stress, enriched in cellular components such as membrane raft and membrane microdomain, and involved in molecular functions such as heme binding and cytokine receptor binding, and were closely related to lipid and atherosclerosis, advanced glycation end products⁃receptor for advanced glycation end products signaling pathway in diabetic complications, etc. Molecular docking results indicated that core compounds such as luteolin, quercetin, and kaempferol could bind well to core target proteins. (2) HE staining results elucidated that after intervention with Bukan Yili Decoction, the esophageal mucosal structure of rats basically returned to normal, the squamous epithelium was repaired intact, the basal cells were arranged in a single layer, and no obvious inflammatory cell infiltration was observed. Compared with the sham operation group, the expressions of JUN and RELA proteins in the tissues from the middle and lower esophagus of rats in the model group were increased (P<0.05). Compared with the model group, the expressions of JUN and RELA proteins in tissues from the middle and lower esophagus of the low⁃, medium⁃, and high⁃dose Bukan Yili Decoction groups were decreased, and the RELA protein expression in tissues from the middle and lower esophagus of the medium⁃ and low⁃dose Bukanyili Decoction groups was lower than that of the high⁃dose Bukan Yili Decoction group (P<0.05). Conclusion Bukan Yili Decoction can exert a therapeutic effect on GERD by downregulating the expressions of target proteins JUN and RELA, and regulating processes such as inflammatory response, oxidative stress, cell apoptosis, and esophageal mucosal barrier repair.

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