Background: Bisphenol A (BPA) is an environmental endocrine-disrupting chemical associated with impaired male reproductive function, oxidative stress, sperm DNA damage, and altered spermatogenesis. Anthocyanins are natural polyphenolic compounds with strong antioxidant and cytoprotective properties, but their role in protecting sperm DNA integrity and spermatogenesis-related gene expression under BPA exposure remains insufficiently clarified.

Objective: This study aimed to evaluate the protective effect of anthocyanins against BPA-induced spermatogenic injury in male rats, with emphasis on epididymal sperm parameters, sperm DNA integrity, chromatin status, and the expression of AKAP4, SYCP3, and TEX11 genes.

Methods: Forty adult male rats were randomly divided into four groups: control (C), BPA-treated (BPA), anthocyanin-treated (ANT), and BPA plus anthocyanin-treated (BPA+ANT) groups. The experiment lasted for 60 days. At the end of the study, epididymal sperm was collected to assess sperm motility, viability, morphology, concentration, and acrosomal integrity. Sperm DNA fragmentation was evaluated using the acridine orange assay, while chromatin maturity and nuclear chromatin condensation were assessed using toluidine blue and aniline blue staining, respectively. Testicular tissue was used for histological examination and quantitative real-time PCR analysis of AKAP4, SYCP3, and TEX11 expression.

Results: BPA exposure significantly reduced sperm motility, viability, normal morphology, concentration, and acrosomal integrity compared with the control group. BPA also markedly increased sperm DNA fragmentation, abnormal nuclear chromatin condensation, and abnormal chromatin maturity. At the molecular level, BPA significantly downregulated AKAP4, SYCP3, and TEX11 expression. Anthocyanin treatment significantly improved sperm functional parameters, reduced DNA fragmentation and chromatin abnormalities, and restored the expression of spermatogenesis-related genes toward control levels in BPA-exposed rats.

Conclusion: Anthocyanins attained BPA-induced spermatogenic injury by preserving sperm quality, sperm DNA integrity, chromatin stability, and the expression of key spermatogenesis-related genes. These findings suggest that anthocyanins may represent a promising natural protective agent against BPA-induced male reproductive toxicity