Screening 16 bisphenol analogues in asthenospermia: BPM-CDK1 axis identified by computational and In Vitro validation

Bisphenol A (BPA) is a ubiquitous industrial chemical found in numerous consumer products. While its endocrine-disrupting effects and reproductive toxicity are well-documented, the widespread use of structural analogues (BPs) as "BPA-free" alternatives has raised urgent concerns regarding their comparable health risks. In this study, we employed a systematic network toxicology framework to evaluate the pathogenic mechanisms of 16 bisphenol congeners in asthenospermia. Through an integrated screening of disease-target associations, five lead compounds-BPA, BPS, BPB, BPP, and BPM-were identified as having the most significant correlation with sperm motility disorders. To elucidate the underlying molecular mechanism, core targets were identified via Protein-Protein Interaction (PPI) networks and further prioritized using LASSO regression, Random Forest (RF), Logistic Regression (LR), Naive Bayes (NB), Support Vector Machine (SVM), and Extreme Gradient Boosting (XGBoost). Molecular docking analysis revealed that CDK1 exhibited the most robust binding affinity to these BPs (lowest docking energy of -10.4 kcal/mol), a finding further validated by molecular dynamics simulations showing high structural stability of the BPM-CDK1 complexes. Notably, among the prioritized congeners, BPM emerged as a critical modulator of this signaling node. In vitro experiments demonstrated that BPM exposure significantly impaired the migration of GC-2spd(ts) and TM4 cells. Furthermore, BPM treatment markedly downregulated CDK1 expression at both the mRNA and protein levels. These results suggest that BPs contribute to the pathogenesis of asthenospermia by disrupting cell proliferation and cycle regulation, specifically through the BPM-mediated suppression of the CDK1 axis.