Astragaloside IV attenuates ferroptosis-related testicular injury in rats with experimental varicocele

Varicocele is an important correctable cause of male infertility. Oxidative stress and lipid peroxidation contribute to the associated testicular injury, but the role of ferroptosis remains controversial. Astragaloside IV (AS-IV), a quality-control metabolite of Astragali radix, has antioxidant activity and can modulate ferroptosis-related processes, yet its effect in varicocele is unclear. This study evaluated whether AS-IV attenuates testicular injury in experimental varicocele and examined ferroptosis-related mechanisms. A Sprague–Dawley rat model was established by partial ligation of the left renal vein and treated with oral AS-IV; Ferrostatin-1 served as a pharmacological comparator. Label-free proteomics, histopathology, biochemical assays and targeted molecular analyses were combined with a TM4 Sertoli-cell model of RSL3-induced injury. AS-IV lessened tubular injury and improved Johnsen scores in a dose-related manner, reduced testicular Fe 2+ , ROS and MDA, and raised the GSH/GSSG ratio. Proteomics showed partial reversal of lipid-metabolism and ferroptosis-related enrichment after AS-IV. These changes were accompanied by recovery of androgen receptor (AR) and MBOAT2. In TM4 cells, AS-IV improved viability and lowered C11-BODIPY oxidation after RSL3; enzalutamide weakened the effects of AS-IV on lipid peroxidation and MBOAT2 and only partly reduced the viability rescue. The findings provide experimental evidence that AS-IV ameliorates varicocele-related testicular injury and that AR/MBOAT2-related signalling may participate.