Abstract: Objective: This study aimed to investigate the effect of blueberry extract on the regulation of the FXR/PI3K/AKT pathway on glycolipid metabolism. Methods: C57BL-6J mice were fed a high-fat diet to induce an obesity model and were fed with blueberry extract (300, 200, 100 mg/kg) for 16 weeks. Comparison of body weight, organ ratios, and serum levels of glucose and lipids in groups of mice. Subsequently, histopathological examination of the liver tissue were completed. Comparison of survival rates, levels of basic glucose consumption, total cholesterol, and triglycerides in groups of cell. The expression of FXR, PI3K, AKT in mouse liver tissue and insulin resistance model cell was detected by Western blot. An insulin resistance model was constructed by inducing BRL-3A in rat hepatocytes with palmitic acid and intervening with blueberry extract (40, 80, 160 μg/mL). Results: Blueberry extract inhibited weight gain and reduced fasting glucose, fructosamine, total cholesterol, triglyceride, high and low-density lipoprotein cholesterol levels in obese mice (P<0.01), and increased the viability of IR cells, cellular basal glucose uptake (P<0.05), and elevated the protein expression levels of FXR, PI3K, and AKT (P<0.05), and showed a dose-effect relationship. Conclusion: Blueberry extract can up-regulate the FXR/PI3K/AKT signaling pathway and alleviate the abnormalities of glycolipid metabolism caused by high-fat exposure.