Copper (Cu) is a widespread environmental heavy metal that plays an essential role as a trace element in organisms. However, growing evidence indicates that excessive Cu exposure has toxic effects on ovarian function. Nevertheless, the underlying mechanisms remain largely unknown. This study aimed to evaluate the toxic effects of Cu on human granulosa cells and explore the potential mechanisms. An exploratory transcriptomic analysis was performed on Cu-exposed cells to investigate identified candidate genes and pathways (based on nominal p < 0.05). Pathway enrichment analysis showed that these DEGs were significantly overrepresented in several pathways, including GSH metabolism, and DNA damage processes. Cu exposure caused a concentration-dependent increase in DNA damage within cells, with oxidative stress serving as the underlying mediator. Disturbed GSH homeostasis contributes to Cu-induced oxidative stress and DNA damage, as verified by pretreatment with GSH‑MEE and BSO. However, without measuring GSH synthesis flux or GSSG, alternative causes of GSH depletion cannot be excluded. The BSO experiment should be interpreted as evidence for GSH depletion and ROS, not as direct confirmation of DNA damage. In summary, these findings elucidate the mechanisms of Cu-induced DNA damage in granulosa cells, thereby providing new perspectives on the pathways underlying female reproductive disorders.
