NEFA Induces Ferroptosis in Transition Dairy Cattle Liver by Increasing Lipid Reactive Oxygen Species and Downregulating SLC7A11

Excessive lipid mobilization in transition dairy cattle leads to elevated levels of nonesterified fatty acids (NEFA), causing liver injury. Ferroptosis, a novel cell death mechanism, has significant roles in liver disease progression. However, its potential role in NEFA-induced liver injury remains unclear. In this study, we demonstrated ferroptosis occurrence in the transition cattle liver through histopathological damage, increased lipid peroxidation, and iron accumulation. In vitro, using ferroptosis inducer RAS-selective lethal 3 (RSL3), NEFA significantly enhanced ferroptosis in bovine hepatocytes. Subsequent experiments with ferroptosis inhibitor ferrostatin-1 (Fer-1) showed Fer-1 markedly reduced NEFA-enhanced ferroptosis, confirming ferroptosis' critical role. Mechanistic studies revealed that NEFA promoted ferroptosis by inducing lipid ROS production effectively inhibited by N-acetylcysteine (NAC) and by downregulating SLC7A11 expression. In conclusion, ferroptosis critically mediates NEFA-induced liver injury in transition dairy cattle, indicating that targeting this pathway may offer potential therapeutic strategies to mitigate liver damage associated with excessive lipid mobilization.