This study shows that overexpression of the G0S2 gene aggravates liver insulin resistance of mice through upregulating P-Foxo1, Socs3, and Ptpn1 and downregulating Gstp1 and Ppar-γ, which demonstrates that G0S2 may be a potential target gene for the treatment of NAFLD, obesity, and diabetes. Here, G0S2 is linked to metabolic dysfunction-associated steatotic liver disease.