Protective effect of high-intensity aerobic training and Crocin on testicular oxidative stress and apoptosis in healthy male Wistar rats treated with doxorubicin

Chemotherapy-induced testicular toxicity remains a significant limitation in male reproductive health, with oxidative stress and apoptosis as key mediators. This study investigated the effects of HIAT and Crocin on doxorubicin (DX)- induced testicular damage. Thirty-five male Wistar rats were allocated into seven groups (n=5): healthy control (HC), doxorubicin control (DXC), DX + Crocin 10 mg/kg (DX-C10), DX + Crocin 50 mg/kg (DX-C50), DX + HIAT, DX + HIAT + C10, and DX + HIAT + C50. Doxorubicin was administered at 2 mg/kg/week for seven weeks. HIAT consisted of 2–8 × 2-min intervals at 80–110% vV̇ O₂max, 5 days/week. Oxidative stress and apoptosis were quantified via MDA levels, and Caspase-3/9 expression (ELISA, qPCR), and testicular morphology was assessed using H&E and DAPI staining. Doxorubicin increased oxidative stress and apoptosis, by elevated MDA and Caspase-3/9 (P < 0.01), and reduced seminiferous tubule diameter (P < 0.01). HIAT, Crocin, and their combination significantly mitigated these effects (P < 0.01). The DX + HIAT + C50 group demonstrated the greatest attenuation of apoptotic markers and histopathological damage, whereas Crocin 50 mg/kg alone reduced MDA levels (P<0.05). These findings suggest that Crocin plus HIAT may serve as a potential strategy to mitigate chemotherapy-related reproductive damage.