Abstract: BACKGROUND: Current therapeutic measures for oligoasthenozoospermia remain limited, while botanical herbal medicines with minimal adverse reactions and abundant sources provide novel insights and approaches for the treatment of oligoasthenozoospermia.
OBJECTIVE: To investigate the mechanism of Allii Tuberosi Semen in improving spermatogenesis and quality in a rat model of oligoasthenozoospermia induced by tripterygium glycoside tablets using network pharmacology, and to conduct experimental verification.
METHODS: This study integrated network pharmacology and bioinformatics approaches to systematically investigate the molecular mechanisms underlying the therapeutic effects of Allii Tuberosi Semen intervention in the rat model of oligoasthenozoospermia induced by tripterygium glycoside. Specifically, active ingredient targets of Allii Tuberosi Semen and oligoasthenozoospermia-associated disease targets were identified through cross-database screening (TCMSP, Genecards, etc.), followed by Venn intersection analysis to pinpoint core therapeutic targets. The drug-component-target-disease network was constructed by Cytoscape, the protein-protein interaction network was constructed in combination with the STRING database, and the top 10 core targets were screened based on topological parameters. DAVID was used to analyze the enrichment of gene ontology function and Kyoto Encyclopedia of Genes and Genomes pathway, revealing that Allii Tuberosi Semen played a therapeutic role by regulating the phosphatidylinositol 3-kinase/protein kinase B (PI3K-AKT) signaling pathway, and CB-Dock2 was used to calculate the kinetic binding energy by molecular docking. Animal experiments were conducted to further verify the mechanism by which Allii Tuberosi Semen improves spermatogenesis and sperm quality in the rat model of oligoasthenozoospermia induced by tripterygium glycoside.
RESULTS AND CONCLUSION: (1) Network pharmacology analysis revealed that Allii Tuberosi Semen contain 16 bioactive compounds, with 1 095 disease-associated targets identified through systematic screening. (2) The protein-protein interaction network screened out 10 core targets. (3) Gene ontology function and Kyoto Encyclopedia of Genes and Genomes pathway analyses demonstrated significant enrichment in the PI3K/AKT signaling pathway. (4) Molecular docking analysis was performed between key targets of the PI3K/AKT signaling pathway (serine kinase 1, B lymphoblastoma 2 protein, interleukin 6, epidermal growth factor receptor, and glycogen synthase kinase 3β) and the bioactive compounds quercetin, kaempferol, and linoleic acid. (5) Animal experiments demonstrated that treatment with Allii Tuberosi Semen significantly improved testicular weight, sperm count, and the proportion of sperm with progressive motility in the rat model of oligoasthenospermia. It effectively ameliorated histopathological manifestations in the testes and modulated the expression of phosphorylated PI3K, phosphorylated AKT, and B lymphoblastoma 2 protein, resulting in a significant reduction in the testicular tissue apoptosis rate. These findings suggest that Allii Tuberosi Semen may improve spermatogenesis and sperm quality in the tripterygium glycoside-induced oligoasthenospermia rat model by regulating the PI3K/AKT pathway.

Key words: Allii Tuberosi Semen, oligoasthenozoospermia, network pharmacology, animal experiment, phosphatidylinositol 3-kinase (PI3K), protein kinase B (AKT), apoptosis, molecular docking, homology of medicine and food