Deoxynivalenol (DON) is a prevalent mycotoxin that causes severe liver damage through hepatocellular oxidative stress. This study investigates the role of the xenobiotic metabolism factor, pregnane X receptor (PXR), in mediating DON-induced hepatocellular oxidative stress. Treatment with the PXR agonist 3-indole-propionic acid (IPA) alleviates DON-induced oxidative stress and liver injury in both in vitro and in vivo models. Mechanistically, IPA directly transactivates the m⁶A demethylase FTO, leading to site-specific demethylation and reduced abundance of YTHDC1-bound Malat1 lncRNA. The decreased m⁶A modification of Malat1 lncRNA reduces its stability and enhances antioxidant pathways governed by NRF2, thereby mitigating DON-induced liver injury. Malat1 knockout mice exhibit decreased DON-induced liver injury, emphasizing the role of Malat1 lncRNA in oxidative stress. These findings establish that PXR-mediated m⁶A-dependent Malat1 lncRNA expression determines hepatocyte oxidative stress via m⁶A demethylase FTO, providing insights into the mechanisms of DON-induced liver injury and potential therapeutic strategies. PXR regulates Malat1 lncRNA expression through post-transcriptional m⁶A modification, which influences NRF2 activity and antioxidant pathways. IPA treatment alleviates DON-induced oxidative stress and liver injury by modulating PXR activity and FTO expression. Malat1 lncRNA, through its interaction with YTHDC1, enhances stability and expression, contributing to oxidative stress. These results highlight the critical role of the PXR-FTO-Malat1 lncRNA-NRF2 pathway in DON-induced liver injury and oxidative stress, offering potential therapeutic strategies for liver damage caused by DON or other xenobiotics.Deoxynivalenol (DON) is a prevalent mycotoxin that causes severe liver damage through hepatocellular oxidative stress. This study investigates the role of the xenobiotic metabolism factor, pregnane X receptor (PXR), in mediating DON-induced hepatocellular oxidative stress. Treatment with the PXR agonist 3-indole-propionic acid (IPA) alleviates DON-induced oxidative stress and liver injury in both in vitro and in vivo models. Mechanistically, IPA directly transactivates the m⁶A demethylase FTO, leading to site-specific demethylation and reduced abundance of YTHDC1-bound Malat1 lncRNA. The decreased m⁶A modification of Malat1 lncRNA reduces its stability and enhances antioxidant pathways governed by NRF2, thereby mitigating DON-induced liver injury. Malat1 knockout mice exhibit decreased DON-induced liver injury, emphasizing the role of Malat1 lncRNA in oxidative stress. These findings establish that PXR-mediated m⁶A-dependent Malat1 lncRNA expression determines hepatocyte oxidative stress via m⁶A demethylase FTO, providing insights into the mechanisms of DON-induced liver injury and potential therapeutic strategies. PXR regulates Malat1 lncRNA expression through post-transcriptional m⁶A modification, which influences NRF2 activity and antioxidant pathways. IPA treatment alleviates DON-induced oxidative stress and liver injury by modulating PXR activity and FTO expression. Malat1 lncRNA, through its interaction with YTHDC1, enhances stability and expression, contributing to oxidative stress. These results highlight the critical role of the PXR-FTO-Malat1 lncRNA-NRF2 pathway in DON-induced liver injury and oxidative stress, offering potential therapeutic strategies for liver damage caused by DON or other xenobiotics.