Curcumin Protects Mouse Spermatogonia from Triptolide-Induced Injury Through Modulation of Ferroptosis-Related Pathways

Triptolide (TP) is an effective anti-inflammatory and immunosuppressive agent, yet its clinical application is constrained by significant male reproductive toxicity. Curcumin, a natural antioxidant, exhibits protective effects; however, whether it protects against TP-induced damage during mouse spermatogenesis and the underlying mechanisms remain incompletely understood. Methods: Proteomic analysis was performed to investigate the protective mechanism of curcumin in mouse GC-1 cells, followed by multiple validation assays including CCK-8 assay, apoptosis detection, measurement of reactive oxygen species (ROS), glutathione (GSH), malondialdehyde (MDA), and Fe2+ levels, quantitative polymerase chain reaction (qPCR), Western blotting (WB), hematoxylin-eosin (HE) staining, and immunofluorescence. Results: Proteomic analysis revealed that curcumin primarily ameliorated TP-induced damage in mouse spermatogonia by modulating ferroptosis-related pathways. Curcumin elevated GSH levels; reduced MDA, ROS, and Fe2+ levels; alleviated lipid peroxidation; and regulated ferroptosis-related pathways in both TP-induced GC-1 cells and testicular tissue. These effects were associated with upregulation of the mRNA and protein expression of Nrf2, Gclc, and Map1lc3a and downregulation of Tfrc and Dmt1. Collectively, these findings demonstrate the protective effect of curcumin against TP-induced spermatogonial damage. Conclusions: Curcumin regulated ferroptosis-related pathways by modulating the expression of Nrf2, Gclc, Map1lc3a, Tfrc, and Dmt1, thereby significantly ameliorating TP-induced damage in mouse spermatogonia.