Ferroptosis Inhibitor Liproxstatin-1 Alleviates Triptolide-Induced Blood Testis-Barrier Disruption and Testicular Injury.

Triptolide (TP), the principal bioactive component of Tripterygium wilfordii (T. wilfordii), exhibits potent pharmacological activities; however, its clinical application is limited by severe male reproductive toxicity, and effective interventions to mitigate reproductive toxicity remain limited. Although the relationship between ferroptosis and TP-induced male reproductive damage has garnered attention, it is still not fully understood whether intervening in ferroptosis can serve as a viable approach to ameliorate TP-induced testicular injury. Thus, this study aimed to characterize TP-induced ferroptosis in mouse testes and Sertoli cells and evaluate whether pharmacological inhibition of ferroptosis by liproxstatin-1 (Lip-1) could serve as a protective strategy against TP-induced testicular injury. In this study, we demonstrated that TP exposure caused significant testicular damage, characterized by pathological histological changes, disruption of the blood-testis barrier (BTB) integrity, a decrease in sperm count and motility, and an increase in sperm deformity. Mechanistically, TP downregulated key redox regulators (NRF2, HO-1, SLC7A11, and GPX4) and iron homeostasis proteins (FTH1, FTL, and FPN), implicating ferroptosis in male reproductive toxicity. Crucially, co-treatment with the Lip-1 effectively mitigated both BTB disruption and testicular damage induced by TP, both in vivo and in vitro. The protective effect was associated with the attenuation of lipid peroxidation and the restoration of the NRF2/HO-1/SLC7A11/GPX4 pathway. Collectively, our findings suggested that pharmacological inhibition of ferroptosis by Lip-1 represented a promising strategy against TP-induced BTB disruption and testicular injury.