Individual and Combined Cytotoxic Effects of Zearalenone and Zearalenone-14-Glucoside in Porcine Intestinal Epithelial Cells.

Zearalenone (ZEN) and its masked form, zearalenone-14-glucoside (Z14G), are common mycotoxins found in feed that can threaten the intestinal health of piglets. To date, there has been a lack of systematic research on the toxic effects and molecular mechanisms of Z14G alone and in combination with ZEN on intestinal porcine epithelial cell-jejunal 2 (IPEC-J2) cells. This study found 80 µmol/L ZEN and 100 µmol/L Z14G were the selected experimental concentrations based on cell viability assays. Both ZEN and Z14G significantly inhibited cell viability and proliferation, induced apoptosis, and disrupted the expression of associated genes. They altered the expression of barrier-associated genes, evidenced by increase in lactate dehydrogenase release and downregulation of tight junction gene mRNA. ZEN and Z14G exposure increased oxidative stress, altered antioxidant defense, and modulated inflammatory responses, accompanied by changes in the expression of genes related to the TLR4/MyD88 and NLRP3 signaling pathways. Notably, Z14G alone exhibited a distinct transcriptional response of inflammatory-related genes, failing to upregulate IL-1β, TNF-α and STING. In conclusion, ZEN and Z14G disrupt intestinal epithelium homeostasis via interlinked oxidative, inflammatory and barrier-damaging pathways. The findings reveal the direct toxicity and unique properties of ZEN and Z14G, underscoring the need to consider masked mycotoxins in combined risk assessment.