Therapeutic effects of Guilu-Erxian-Glue treatment on oligoasthenospermia: Evidence from network pharmacology, molecular docking, and in vivo experimental validation

•We have constructed a network of disease-drug-compound-target and identified therapeutic candidate targets for GLEXG against OAS.•Molecular docking revealed that hub targets screened in the PPI network showed a tight binding with potential bioactive compounds in GLEXG.•An OAS rat model was established to further verify the therapeutic effects of GLEXG. Oligoasthenospermia (OAS) remains the leading cause of male infertility. Guilu-Erxian-Glue (GLEXG) is a traditional Chinese formula that exhibits beneficial effects in attenuating reproductive dysfunction. However, the mechanism of GLEXG against OAS remains unclear. In this study, we investigated the therapeutic effects of GLEXG against OAS and analyzed its constituents and underlying mechanism. Using the network pharmacology analysis, 40 targets and 53 bioactive compounds of GLXEG associated with OAS were identified, forming a tight disease-drug-compound-target network. IL-6, MYC, CASP3, EGFR, VEGFA, ERBB2, and ESR1 were screened as hub targets in the protein-protein-interaction network. KEGG and GO enrichment analysis and pathway-target-compound network indicated that the HIF-1 signaling pathways were enriched by numerous targets and interact with multiple pathways, including PI3K/Akt and TNF pathway. Molecular docking analysis revealed a tight binding between potential bioactive compounds and hub targets, and the molecular dynamic simulation of the ESR1-Frutinone A complex with the lowest docking energy exhibited high stability. In addition, an OAS rat model was established to determine the therapeutic effects. The results indicated that GLEXG treatment markedly improved sperm quality in the epididymis and reversed histopathological lesions in the testicular tissue, and regulated the expressions of the predicted hub targets and HIF-1α associated with HIF-1 signaling pathway. Our results illustrated the potential therapeutic effects of GLEXG on OAS, suggesting that it is an alternative therapy against OAS.