Melphalan’s Effect on Testicular Prepubertal Rats: Induction of Autophagy and Apoptosis, Modulated by the Potential of Chrysin

Melphalan, a widely used alkylating chemotherapeutic agent, is known to cause gonadotoxicity, particularly affecting spermatogenesis and steroidogenesis. This study investigated the protective effects of chrysin, a flavonoid with antioxidant and antiapoptotic properties, on melphalan-induced testicular damage in immature male rats. Forty-eight prepubertal Wistar rats were divided into six groups receiving melphalan (0.5 mg/kg/day, intraperitoneally), chrysin (50 or 75 mg/kg/day, orally), or combinations thereof. Treatment spanned 30 days, covering the prepubertal to pubertal transition. Melphalan exposure significantly reduced testosterone levels, disrupted spermatogenesis, and increased oxidative stress, as indicated by elevated malondialdehyde and decreased glutathione peroxidase levels. Gene expression analysis revealed marked upregulation of autophagy (LC3β, Beclin-1, Atg5, Atg7, Atg12) and apoptosis (BAX, caspase-3) markers, alongside downregulation of the antiapoptotic gene BCL2. Histological analysis showed severe damage to seminiferous tubules and reduced Johnsen scores. Chrysin coadministration, particularly at 75 mg/kg, significantly ameliorated these effects by restoring antioxidant capacity, reducing expression of cell death-related genes, and improving testicular structure and hormone levels. These findings demonstrate that chrysin mitigates melphalan-induced gonadotoxicity through modulation of oxidative stress, apoptosis, and autophagy pathways. Chrysin's dose-dependent protective effects highlight its potential as a natural therapeutic agent to preserve reproductive function in individuals undergoing chemotherapy during prepuberty.