Salidroside mitigates Oligoasthenospermia by attenuating ferroptosis via NF-κB pathway inhibition

BACKGROUND: Oligoasthenospermia (OAS) leads to male infertility, significantly impacting patients and their families. Previous studies have identified various mechanisms involved in OAS, including oxidative stress, apoptosis, and autophagy. Ferroptosis, a novel cell death form, is also linked to male infertility. Despite advancements, the involvement of ferroptosis in spermatogenesis and its potential role in causing OAS, as well as the identification of safe and effective natural drugs targeting this mechanism, remain unclear. PURPOSE: This study aimed to investigate the potential role of ferroptosis in OAS and explore safe and effective inhibitors for treatment. STUDY DESIGN AND METHODS: Semen samples were collected from patients with OAS. Mouse models of OAS were induced using cyclophosphamide (CP) in vivo, and GC-2 cells were treated in vitro. The effects of salidroside (Sal) on mice and GC-2 cells were evaluated using various methods, including ATP, malondialdehyde, and glutathione peroxidase activity assays; transmission electron microscopy; immunohistochemistry; flow cytometry; lipid peroxidation (LPO) analysis; mitochondrial membrane potential (MMP) assessment; enzyme-linked immunosorbent assays; qRT-PCR; western blotting; RNA sequencing; and immunofluorescence. RESULTS: Human OAS sperm exhibits biochemical and morphological hallmarks of ferroptosis. Ferroptosis is crucial in CP-induced OAS in mice in vivo, and its inhibition can effectively improve OAS. Further search for safe and stable natural compounds found Sal improved the sperm quality by attenuating ferroptosis-mediated LPO in germ cells through increased GPX4 expression, thereby ameliorating OAS in vivo. Immunohistochemistry revealed that Sal increases GPX4 expression starting from primary spermatocytes, highlighting its action stage during spermatogenesis. In vitro, these results were verified using a mouse spermatocyte cell line, GC-2 cells. Mechanistically, RNA-Seq and bioinformatic analysis suggest that Sal likely promotes GPX4 expression by inhibiting the NF-κB pathway, thereby reducing ferroptosis. CONCLUSION: Ferroptosis plays an important role in OAS, and Sal could ameliorate OAS by attenuating ferroptosis via NF-κB pathway inhibition.