Effects of Zearalenone on Apoptosis and Copper Accumulation of Goat Granulosa Cells In Vitro

SIMPLE SUMMARY: Zearalenone (ZEA) is a prominent feed contaminant worldwide, causing considerable financial losses in the livestock industry due to its estrogen-like effect. Although many reports have claimed that ZEA can interfere with the reproductive system of mammals, little confirmatory information is available. We used ZEA to treat goat granulosa cells (GCs) and found that ZEA could promote apoptosis and oxidative stress levels in GCs, inhibit cell proliferation and arrest the cell cycle. RNA-Seq was performed, which also showed that copper transport-related gene expression was altered, leading to copper accumulation in GCs. These findings provide a new perspective for research on the relationship between ZEA toxicity and copper accumulation ABSTRACT: Zearalenone (ZEA), also known as F-2 toxin, is a mycotoxin. Despite numerous reports of ZEA impairing livestock production performance and fertility, little information is available, including information about the mechanism underlying damage to cell metal ion transport. Copper, which is essential for cell survival as a metal ion, can consist of a variety of enzymes that facilitate abundant metabolic processes. However, the accumulation of copper in cells can have toxic effects. Here, we intended to determine whether ZEA could impair goat granulosa cells (GCs) and alter the cellular copper concentration. GCs were divided into a negative control (NC) group (cells cultured with 0.1% dimethyl sulfoxide (DMSO) for 8 h) and a ZEA group (cells cultured with 200 μmol/L ZEA diluted in DMSO for 8 h). The results showed that ZEA could inhibit GC proliferation and impair cell viability. GCs showed significant increases in the apoptosis rate and oxidative stress levels, while their ability to synthesize estrogen decreased. In addition, RNA-seq results showed dramatic changes in the expression of copper transport-related genes. The expression levels of ATPase copper transporting alpha (ATP7A) and ATPase copper transporting beta (ATP7B) were significantly downregulated (p < 0.01), while the expression of solute carrier family 31 member 1 (SLC31A1) was not modified in the ZEA group compared with the NC group. In accordance with these trends, the copper concentration increased significantly in the ZEA group (p < 0.01). In summary, our results show that ZEA can negatively affect GCs and cause copper accumulation. This finding may provide a prospective line of research on the relationship between ZEA and the transport of copper ions in GCs.