Specnuezhenide attenuates bisphenol A-induced testicular damage through inhibiting iron accumulation, ferroptosis and apoptosis in mice

Background: Bisphenol A (BPA), a xenoestrogenic compound, disrupts spermatogenesis by inducing oxidative stress (OS) through iron ion-dependent mechanisms, ultimately contributing to male infertility. Specnuezhenide (SPN), a secoiridoid derived from Fructus Ligustri Lucidi (FLL) , exhibits notable antioxidant and anti-inflammatory properties. However, whether SPN can protect against BPA-induced OS and its detrimental effects on spermatogenesis remains unclear. Furthermore, the underlying mechanisms by which SPN alleviates BPA-induced male reproductive toxicity are poorly understood. Purpose: To demonstrate the efficacy of SPN in mitigating BPA-provoked testicular damage. Study Design : Specnuezhenide was verified to attenuates bisphenol A-induced testicular damage through inhibiting iron accumulation, ferroptosis and apoptosis in mice. Methods: The male ICR mice have been divided into five groups to investigate these questions, including: the control group, the BPA group (50 mg/kg [bw], orally for 28 days), and three SPN+BPA groups receiving BPA (50 mg/kg [bw], orally for 28 days) along with SPN (30 mg/kg [bw], orally for 21, 28, and 35 days, respectively). The extent od testicular damage was evaluated by basic parameters of body weight, sperm quality, hormonal levels and hematoxylin-eosin (H&E) staining. The mRNA and protein levels of ferroptosis and apoptosis pathways in testes were evaluated by qPCR amplification, western blotting and immunofluorescence analysis. Results: BPA exposure significantly impaired sperm quality, induced OS, caused iron accumulation, and led to mitochondrial damage; restored serum hormone levels, including testosterone (T), luteinizing hormone (LH) and follicle stimulating hormone (FSH), while increasing estradiol (E2) levels; reduced the activities of antioxidant enzyme, such as superoxide dismutase, catalase, and glutathione peroxidase; and sharply elevates in the expressions of NCOA4 ( a marker of ferritinophagy), GPX4 and SLC7A11 (markers of ferroptosis), cysteine- dependent aspartate-specific protease-3 (Caspase-3), cysteine- dependent aspartate-specific protease-9 (Caspase-9) and BCL2-associated X protein (Bax) (markers of apoptosis). Conversely, SPN supplementation considerably mitigated BPA-induced testicular damage by inhibiting iron accumulation and OS, thereby downregulating ferroptosis and apoptosis pathways. Conclusions: These findings underscore potential of SPN as a therapeutic agent and highlight the necessity for in-depth investigation into the detailed mechanisms underlying BPA-induced toxicity.