Integrated Network Pharmacology, Molecular Docking, and Nanoformulation Approach to Elucidate the Therapeutic Potential of Fisetin in Male Infertility

Abstract

Background: Male infertility is frequently associated with urogenital bacterial infections and oxidative stress, which impair sperm function and reproductive health. Natural flavonoids such as fisetin have gained attention due to their antimicrobial, antioxidant, and anti-inflammatory properties.

Objective: The study aimed to investigate the therapeutic potential of fisetin in male infertility using an integrated approach combining network pharmacology, molecular docking, nanoformulation, and antimicrobial evaluation.

Methods: Potential targets of fisetin were predicted using in silico tools and intersected with male infertility-associated genes to identify common targets. Protein–protein interaction (PPI) analysis, Gene Ontology (GO), and KEGG pathway enrichment were performed to determine key biological processes and pathways. Molecular docking using AutoDock evaluated binding interactions of fisetin with hub proteins. Fisetin-loaded liposomes were prepared by the thin-film hydration method and characterised for particle size, polydispersity index, zeta potential, drug loading, and morphology. Antimicrobial activity was assessed using the minimum inhibitory concentration (MIC) assay against Staphylococcus aureus and Escherichia coli.

Results: A total of 32 common targets were identified, with hub genes including AKT1, PIK3R1, EGFR, IGF1R, INSR, SRC, and MET. Enrichment analysis revealed the PI3K–Akt and MAPK signalling pathways as key regulators in male infertility. Molecular docking showed strong binding affinity of fisetin towards major targets. The liposomal formulation exhibited a particle size of 153.26 nm, low PDI (0.136), and zeta potential of −24.74 mV, indicating good stability. Fisetin demonstrated antimicrobial activity with MIC values of 2.5 mg/mL (S. aureus) and 1.25 mg/mL (E. coli).

Conclusion: Fisetin exerts therapeutic effects through a multi-target mechanism, modulating key signalling pathways involved in oxidative stress, inflammation, and cellular survival. Liposomal encapsulation enhances its physicochemical properties, while its antimicrobial activity may contribute to managing infection-associated infertility. These findings support fisetin as a promising candidate for further in vivo and clinical evaluation.