Ganoderma lucidum mitigates oxidative stress and apoptosis in chlorpyrifos‑induced testicular toxicity in male rats

Chlorpyrifos (CPS) is an organophosphate pesticide known to induce oxidative stress, apoptosis, and histopathological damage in male reproductive tissues. Ganoderma lucidum (GDL), a medicinal fungus rich in bioactive polysaccharides and triterpenoids, has demonstrated potent antioxidant and antiapoptotic properties in various models. This study aimed to evaluate the protective effects of GDL extract against CPS-induced testicular toxicity in male rats by assessing histological integrity, oxidative stress markers, apoptosis signaling, and hormonal balance. Twenty-four adult male Wistar rats were randomly divided into four groups (n = 6): Cont, CPS (20 mg/kg), GDL (150 mg/kg), and Treat. Treatments were administered orally once daily for 30 days. At the end of the study, the testes were harvested for morphometric and Cosentino histopathological scoring, immunohistochemistry for BAX, BCL2, and Ki67, RTqPCR quantification, biochemical assays of antioxidant enzymes and serum testosterone, LH, and FSH measurements. CPS treatment significantly elevated histopathological damage scores, and disrupted regular cell arrangement. IHC revealed increased BAX and decreased BCL2 and Ki67 expression, corroborated by the upregulation of Bax and Caspase3 transcripts and downregulation of Bcl2. Antioxidant enzymes were suppressed and MDA was elevated in the CPS group. Hormonal assays showed decreased testosterone and LH. Co-administration of GDL partially restored histological architecture, normalized Voronoi patterns, attenuated apoptotic marker alterations, enhanced antioxidant defenses, reduced lipid peroxidation, and recovered hormone levels toward control values. GDL extract confers significant protection against CPS-induced testicular injury by mitigating oxidative stress, modulating apoptotic pathways, and preserving endocrine function, highlighting its therapeutic potential against pesticide-related reproductive toxicity.