ArticleScientific reports2025
The therapeutic potential of Zuogui Wan in oligoasthenozoospermia: insights from network pharmacology, molecular docking, molecular dynamics simulation, and experimental validation.
Article in Scientific reports, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
Oligoasthenozoospermia (OAS) is a major cause of male infertility, with limited effective treatments. Chinese patent medicine Zuogui Wan (ZGW) has been traditionally used to improve sperm quality, but its molecular mechanisms remain unclear. This study integrates network pharmacology, molecular docking, molecular dynamics (MD) simulation, and in vivo and in vitro experiments to explore ZGW's therapeutic effects in OAS. Active compounds and targets of ZGW were identified using network pharmacology, and intersecting OAS-related targets underwent enrichment and protein-protein interaction (PPI) analysis. Molecular docking and MD simulations assessed compound-target binding affinity and stability. In vitro, CCK-8 assays measured cell proliferation, while qPCR and Western blot analyzed key gene and protein expression. In vivo, a rat OAS model was used to evaluate ZGW's therapeutic effects through transmission electron microscopy (TEM), hematoxylin & eosin (HE) staining, and TUNEL assays. The expression of key molecular targets was further validated by qPCR and Western blot. A total of 182 potential targets were identified, with TP53, NF-κB1, and PKC as key hub genes. KEGG pathway analysis highlighted the involvement of the PI3K-AKT and MAPK signaling pathways.Four core bioactive compounds-Cyasterone, Betavulgarin, Kaempferol, and Quercetin-were identified, with Cyasterone exhibiting the strongest binding affinity and highest stability.In vitro experiments demonstrated that ZGW significantly promoted cell proliferation and regulated apoptosis-related gene expression, indicating its potential in enhancing sperm function. In vivo, ZGW improved testicular structure, enhanced sperm quality, and reduced spermatogenic cell apoptosis, as evidenced by TEM, HE, and TUNEL assays. Molecular validation further confirmed ZGW's modulation of key signaling pathways involved in OAS. ZGW modulates apoptosis, oxidative stress, and key pathways (PI3K-AKT, MAPK) while regulating TP53, NF-κB, and PKC expression. Cyasterone exhibits strong binding and stability with core targets. This study supports ZGW as a potential treatment for male infertility.
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