Protective and therapeutic roles of Withania Somnifera in cyclophosphamide-induced ovarian dysfunction in rats

Background Chemotherapy-induced ovarian dysfunction is a major cause of premature ovarian insufficiency (POI), infertility, and follicular depletion. Cyclophosphamide (CP), a widely used alkylating chemotherapeutic agent, exerts pronounced gonadotoxic effects through DNA damage, oxidative and nitrosative stress, and inflammatory signaling. Objective To evaluate the protective and therapeutic effects of Withania somnifera (Ashwagandha) root extract against CP-induced ovarian dysfunction in rats. Methods Thirty-six mature female albino rats were randomly allocated into six groups: negative control, CP-treated groups (protective and therapeutic timing), W. somnifera -treated group, and combined CP + W. somnifera groups (protective and therapeutic paradigms). Ovarian dysfunction was induced by a single intraperitoneal dose of CP (200 mg/kg). Ovarian histopathology, oxidative and nitrosative stress markers, serum reproductive hormones, DNA damage using alkaline and neutral comet assays, and ovarian expression of the pro-inflammatory cytokines (TNF-α and TGF-β) were assessed. Results CP administration induced marked ovarian injury, as evidenced by significant DNA damage, upregulation of TNF-α and TGF-β, disruption of gonadotropin-estradiol balance, elevated oxidative and nitrosative stress, and pronounced histopathological degeneration. Administration of W. somnifera , particularly in the protective regimen, significantly attenuated CP-induced DNA fragmentation, reduced oxidative and inflammatory markers, partially normalized hormonal profiles, and preserved ovarian histoarchitecture compared with CP-only groups. Overall, the protective regimen showed greater efficacy than the therapeutic regimen. Conclusion W. somnifera exerted significant protective and therapeutic effects against CP-induced ovarian toxicity, primarily through antioxidant, anti-inflammatory, and genoprotective mechanisms. These findings support further investigation of W. somnifera as a potential adjunctive strategy to mitigate chemotherapy-associated ovarian toxicity.