Effect of spermidine on cuproptosis in follicular granulosa cells

1. A study was conducted to investigate the effect of spermidine on cuproptosis in granulosa cells of goose ovarian follicles. Granulosa cells from F2-F5 grade follicles of Sichuan white geese were isolated and cultured.2. Copper sulphate, spermidine and chloroquine were administered to granulosa cells (GC). The GC activity, intracellular Cu²⁺ content, antioxidant enzyme activity, expression levels of genes and proteins related to autophagy and cuproptosis were determined.3. The results showed that spermidine significantly decreased reactive oxygen species (ROS) levels, mitochondrial membrane potential and Cu²⁺content (p < 0.05), and significantly increased the granulosa cell activity (p < 0.001). At the same time, spermidine significantly decreased the expression levels of ferredoxin 1 (FDX1) gene (p < 0.001), lipoylated-dihydrolipoamide S-acetyltransferase (Lip-DLAT) protein (p < 0.05), and increased the expression levels of LC3 gene (p < 0.001). After the cells were treated with spermidine mixed with chloroquine for 8 h, the cells were treated with copper sulphate, and GC activity was significantly decreased (p < 0.001). The expression levels of p62 and FDX1 genes were significantly increased (p < 0.01) and protein levels of LC3 and p62 were significantly increased (p < 0.05).4. In summary, spermidine reduced ROS accumulation, improved mitochondrial membrane potential and increased granulosa cell activity. It alleviated copper sulphate-induced lipoylated protein aggregation and cuproptosis in GC by inducing autophagy. This suggested that spermidine has a broad application prospect in alleviating cuproptosis in GC.