In this study, the participation of glycogen synthase kinase-3β (GSK-3β) in the lipid deposition was investigated in the liver of large yellow croaker (Larmichthys crocea) by LiCl treatment. It was found that the expression of GSK-3β and peroxisome proliferator-activated receptor-γ (PPARγ) was inhibited, but the expression of β-catenin was induced by LiCl treatment. Furthermore, the gene expression and activity of fatty acid synthetase (FAS) and lipoprotein lipase (LPL) in the liver was inhibited by LiCl treatment. The content of total cholesterol (TC), triglyceride (TG), and non-estesterified fatty acid in the liver, as well as TC, TG, and low-density lipoprotein cholesterol in plasma, was decreased by LiCl treatment. However, high-density lipoprotein cholesterol in plasma was increased, and the number of lipid droplets in the liver was decreased by LiCl treatment. The results indicate that GSK-3β/β-catenin may participate in regulating LPL and FAS through PPARγ in the liver of large yellow croaker, which will lead to the inhibition of hepatic lipid deposition.