Salidroside attenuates busulfan-induced testicular injury via activation of The Nrf2/GPX4 axis in mice.

The use of drugs such as busulfan (BUS) often leads to male infertility. Testicular injury represents a significant etiology of male infertility. Ferroptosis has been proven to contribute to testicular impairment. However, clinically effective pharmacological interventions for testicular injury remain limited. The development of novel therapeutics is therefore imperative. Naturally derived bioactive compounds exhibit promising medicinal properties, among which salidroside (SAL) demonstrates multifaceted biological and pharmacological activities. This study aims to investigate ferroptosis involvement in BUS-induced testicular injury and evaluate the therapeutic potential of SAL. Mice were treated with SAL at 25, 50, or 100 mg/kg. Computer-assisted sperm analysis, hematoxylin-eosin staining showed 50 mg/kg SAL yielded the optimal efficacy in improving oligoasthenospermia and repairing testicular/epididymal tissue damage. Further experiments confirmed SAL enhanced the blood-testicular barrier, restored sperm cytoskeleton and normalized abnormal serum hormone levels. Network pharmacology, Western Blot and immunofluorescence revealed SAL regulates the Nrf2/GPX4 axis to inhibit ferroptosis and alleviate oxidative stress. In conclusion, SAL promotes the reversal of BUS-induced testicular injury by mitigating oxidative stress and inhibiting ferroptosis.