Choline reduces inflammatory response and lipid accumulation in calf hepatocytes induced by NEFA via inhibiting ROS/ NF-κB/NLRP3 inflammasome axis

Perinatal dairy cows are highly susceptible to fatty liver syndrome, a prevalent metabolic disorder characterized by excessive hepatic lipid accumulation. Although inflammatory progression drives the transition from physiological lipid storage to pathological steatosis, the underlying mechanisms remain incompletely understood. This study investigates the therapeutic potential of choline in non-esterified fatty acid (NEFA)-induced hepatic inflammation and lipid metabolism dysregulation using an in vitro calf hepatocytes model. A hepatic steatosis model was established by treating primary calf hepatocytes with 1.2 mM NEFA for 24 h. Choline supplementation (75 μM) significantly enhanced hepatocyte viability and reduced intracellular triglyceride (TG) content (P < 0.05). Transcriptomic analysis demonstrated that choline downregulated key components of the NLRP3 inflammasome (P < 0.05) and interleukin-1β (IL-1β) expression (P < 0.05). Mechanistically, choline attenuated NF-κB phosphorylation (P < 0.05) and suppressed intracellular reactive oxygen species (ROS) generation (P < 0.05), thereby inhibiting NLRP3 inflammasome assembly and subsequent IL-1β maturation (P < 0.05). Notably, activation of NLRP3 inflammasome complete abolished choline's protective effects on lipid homeostasis and inflammatory responses (P < 0.05). These findings demonstrate that choline ameliorates NEFA-induced hepatic steatosis by modulating the ROS/NF-κB-NLRP3-IL-1β/IL-18 signalling axis, providing novel therapeutic targets for intervention of steatohepatitis in periparturient dairy cows.