Abstract: Objective: This study aimed to investigate the efficacy and mechanisms by which Chuju flavonoids ameliorated lipid metabolism disorders induced by a high-fat diet in mice, thereby providing a theoretical foundation for dietary interventions in nonalcoholic fatty liver disease (NAFLD). Methods: Seventy male C57BL/6 mice were acclimatized for one week and subsequently assigned to the following groups: normal diet (ND), high-fat diet with low-dose Chuju flavonoid-rich extracts (CJE) (CJEL), positive control (CJE), model group (HFD), and high-fat diet with high-dose CJE (CJEH). Modeling and administration occurred concurrently over a period of 14 weeks. Measurements included body weight, organ indices, and serum and hepatic lipid parameters. The effects of CJE on lipid metabolism disorders induced by a high-fat diet were evaluated through histopathological staining, gut microbiota 16S rRNA sequencing, and Western blot analysis. Results: High-dose Chuju flavonoids significantly decreased total cholesterol (39%±3.8%), triglycerides (41.85%±3.75%), and non-esterified free fatty acids (31.1%±2.4%) in the livers of mice subjected to a high-fat diet. Additionally, these flavonoids inhibited the activity of alanine aminotransferase (32.4%) and aspartate aminotransferase (29.8%). Histopathological analysis demonstrated that CJE markedly reduced hepatic lipid accumulation, as evidenced by a 62.5%±6.2% decrease in lipid droplet area in NAFLD mice. Furthermore, 16S rRNA sequencing confirmed that CJE altered the gut microbiota and activated the AMPK/PPAR-α signaling pathway, which inhibited lipid synthesis and enhances fatty acid β-oxidation. Conclusion: CJE significantly ameliorates lipid metabolism abnormalities in NAFLD mice by restoring gut microbiota balance, activating the AMPK/PPAR-α pathway to suppress hepatic lipid synthesis, and promoting hepatic fatty acid oxidation. This approach offers a viable dietary intervention strategy for the prevention of NAFLD.