Using both constitutive and induced knockouts of Hmgb1 gene selectively in hepatocytes, we demonstrated that HMGB1, acting in the nucleus, exerts a potent repressive effect on LXRα and PPARγ activities and hepatic lipogenesis during metabolic stresses, such as F/R or HFD feeding, suggesting a protective role on the development of NAFLD. This evidence concerns the gene HMGB1 and metabolic dysfunction-associated steatotic liver disease.