Ginsenoside Rc Improves Chilled Storage Quality of Lower-Layer X-Bearing Dairy Goat Spermatozoa Through Enhanced Mitochondrial Function and Antioxidant Capacity

Sex-control technologies that alter offspring sex ratios can markedly improve the economic efficiency of dairy production. Recent studies have demonstrated that treatment with the TLR7/8 agonist R848 enables successful in vitro separation of X- and Y-bearing spermatozoa in mice, cattle, and goats. This is because high-quality semen is essential for successful artificial insemination, especially during chilled storage. In this study, X/Y sperm-specific activation and sorting methods were used to isolate sperm from dairy goats, and the effectiveness of sperm isolation was evaluated using TLR7 immunofluorescence staining. Different concentrations of ginsenoside Rc were added to the semen diluent as a natural preservative, and the lower-layer X-bearing sperm recovered from the sorting system were subjected to chilled storage. The effects of ginsenoside Rc on chilled storage performance were then assessed. Among the tested concentrations, 150 μg/mL of ginsenoside Rc exhibited the most potent protective effect, significantly improving the quality, mitochondrial function, antioxidant capacity and the relative protein abundance of SIRT1, PGC-1α, and Nrf1 of the lower-layer X-bearing sperm after chilled storage for 7 days. In summary, supplementation with ginsenoside Rc in the chilled extender improved the quality of lower-layer X-bearing dairy goat spermatozoa during chilled storage by maintaining mitochondrial function, enhancing antioxidant defense, and reducing oxidative damage, facilitating the application of lower-layer X-bearing sperm-selected sperm in production.