In this study, we found that high dietary fructose consumption promotes MASLD development and progression in C57BL/6J mice, identified Usp2 as a specific fructose-responsive gene, and revealed that the USP2-mediated C/EBPα/11β-HSD1 signaling is involved in disrupting cortisol homeostasis. Here, USP2 is linked to metabolic dysfunction-associated steatotic liver disease.