<p> This study identifies the role of stigmasterol in preserving male fertility via mitigating testicular and sperm destruction in high oxidative stress condition. Methods: Oxidative stress was induced in male mice via oral administration of 2% hydrogen peroxide (H₂O₂) for seven consecutive days, followed by administration of stigmasterol (10 and 20 mg/kg b.w.) orally for another seven days. At the end of the experiment, the mice were sacrificed, then blood, testes, and cauda epididymal sperm were immediately harvested. Sperm parameters, testicular morphology, levels of serum reproductive hormones and expression of proteins (MDA, SOD, CAT, GPx, Keap-1, Nrf-2, HO-1, NqO-1 – markers for oxidative stress), (StAR – marker for steroidogenesis) and (ZO-2 and Connexin-43 - markers for blood-testes barrier integrity) were evaluated. Results: Stigmasterol restores testicular morphology, serum testosterone, LH and FSH levels, sperm parameters and Nrf2, SOD, CAT, GPx, Nqo1, HO-1, StAR, ZO-2, and Connexin-43 expression levels while decreases MDA and Keap1 expression levels in mouse testes.Stigmasterol also help to restore sperm motility, viability, its membrane integrity, and Nqo1 antioxidant levels and decreases sperm DNA fragmentation. Conclusion: Stigmasterol could be used to restore male fertility following exposure to high oxidative stress condition.</p>