Zearalenone toxin induces pyroptosis by activating mitochondrial DNA-STING-NFκB axis in testis and TM4 cell damage

Zearalenone (ZEA), a common mycotoxin in grains and co-products, poses significant reproductive toxicity. However, the toxicity mechanism between ZEA and reproductive toxicity remains unclear. Mitochondrial DNA (mt DNA) alteration is considered to be an important factor in sterility. However, the role of mt DNA in ZEA-induced male reproductive toxicity remains unclear. In this study, we aimed to explore the toxicity effects and mechanism of ZEA in male BALB/c mice and TM4 cells. The mice were treated with 20 mg/kg ZEA 30 d, and cells were treated with 0-20 μmol/L ZEA 24 h. We confirmed that ZEA caused testicular and sperm damage in mice, led to oxidative stress and inflammation in testis, and damaged the blood-testis barrier. In vitro, ZEA caused inflammation and pyroptosis in TM4 cells. In addition, ZEA caused mitochondrial damage, released mt DNA to cytoplasm, and activated STING/NFκB pathway. Interestingly, inhibiting the activation of mt DNA/STING/NFκB alleviated the damage in TM4 cells caused by ZEA. In conclusion, our research has confirmed that ZEA caused testis damage by mt DNA/STING/NFκB pathway. This highlighted the critical role of mt DNA in male reproductive health and provided promising insights into protective mechanisms against environmental toxin-induced male reproductive disorders.