Multi-target protective effects of Pterostilbene against BPA-induced reproductive toxicity revealed by integrative network pharmacology and in vivo validation.

Pterostilbene (PT) is a naturally occurring dimethylated analogue of resveratrol with improved lipophilicity and metabolic stability, yet its role in male reproductive toxicity remains largely unexplored. Using an adult male zebrafish model, this study investigated the protective effects of PT against bisphenol A (BPA)-induced reproductive dysfunction. Adult male zebrafish were randomly assigned to five groups: control, BPA, BPA + PT-L (5 mg/kg), BPA + PT-H (10 mg/kg), and BPA + icariin (ICA, 10 mg/kg). Exposure to BPA (3000 μg/L) resulted in pronounced testicular damage, disrupted spermatogenesis, decreased testosterone levels, and elevated oxidative stress. Administration of PT (5 or 10 mg/kg) markedly improved gonadal morphology and function, restored gonadosomatic index and testosterone levels, and enhanced sperm quality and DNA integrity. Additionally, PT treatment attenuated oxidative damage by reducing reactive oxygen species and lipid peroxidation and by re-establishing endogenous antioxidant capacity. Integrated analyses combining network pharmacology, molecular docking, and gene expression validation indicated that PT acts through multiple targets and pathways associated with cell survival, oxidative stress regulation, and endocrine signaling, including PI3K/Akt, mTOR, MAPK/FoxO, and estrogen-related pathways. These findings suggest that PT may mitigate BPA-induced male reproductive toxicity through coordinated redox and endocrine regulatory mechanisms.