Guilu Erxian glue mitigates spermatogenesis dysfunction through HIF-1α/SLC7A11-mediated ferroptosis inhibition: An integrated metabolomics and network pharmacology study.

BACKGROUND

Oligoasthenospermia is emerging as a critical cause of male infertility resulting from spermatogenesis dysfunction (SGD). Guilu Erxian glue (GLEXG) has traditionally been used to improve sperm quality, but its mechanism remains unclear.

PURPOSE: This study investigates the therapeutic mechanism of GLEXG and its active ingredient quercetin in a Tripterygium wilfordii polyglycoside (GTW)-induced SGD mouse model and GC-1 spermatogonial cells.

METHODS

An SGD model was established by administering GTW (60 mg/kg/day, oral gavage) to BALB/c mice for four weeks, followed by treatment with GLEXG (2.25, 4.50, or 9.00 g/kg/day) or vitamin E (0.02 g/kg/day) for another four weeks. Therapeutic effects were assessed by sperm parameters. Key bioactive constituents and mechanistic pathways were identified using integrated network pharmacology and metabolomics analyses. The role of ferroptosis and associated signaling pathways was validated.

RESULTS

GLEXG restored sperm motility to 78 % and sperm concentration to 67 % of normal levels (p < 0.05) in SGD mice and improved testicular histopathology. Metabolomics indicated protection against ferroptosis through modulation of glutathione metabolism. Quercetin was identified as the key component targeting hypoxia-inducible factor-1alpha (HIF-1α). In erastin-induced ferroptosis, GLEXG-containing serum and quercetin restored GC-1 cell viability by 60 % and 46 %, respectively; reduced lactate dehydrogenase release (76 %; 50 %), reactive oxygen species (ROS) (67 %; 53 %), malondialdehyde (MDA) (72 %; 54 %), and Fe²⁺ (96 %; 86 %); elevated the glutathione/glutathione disulfide (GSH/GSSG) ratio (78 %; 54 %); downregulated HIF-1α (64 %; 45 %); and upregulated glutathione peroxidase 4 (GPX4) (63 %; 32 %) and solute carrier family 7 member 11 (SLC7A11) (64 %; 42 %) (p < 0.05). These effects were reversed by HIF-1α overexpression. In vivo, HIF-1α overexpression abrogated quercetin's protection on sperm motility (53 %), sperm concentration (53 %), and testicular lesions (p < 0.05).

CONCLUSION

GLEXG and quercetin alleviate GTW-induced SGD by inhibiting ferroptosis via HIF-1α/SLC7A11.