Abstract: Objective: To investigate the improvement effect of total flavonoids of Chuju (Chrysanthemum morifolium) on hypercholesterolemia induced by a high-fat diet in mice and explore its mechanism through network pharmacology. Methods: A well-established mouse model of hypercholesterolemia was established using a high-fat diet, administered intragastrically at low, medium or high (50, 100, 200 mg/kg) doses, together with normal control group, milk thistle group and model group. After completing the feeding interventions, appropriately related blood biochemical indexes were analyzed, and the liver was harvested for pathological examination. The results of these investigations were combined with network pharmacological experiments, and key targets and pathways of total flavonoids of Chuju in improving hypercholesterolemia were preliminarily screened, and its possible mechanism was predicted. Results: Compared with the model group, the levels of TC, LDL-C and MDA decreased significantly, ALT and AST activities decreased significantly, and the activities of SOD and GSH-Px increased significantly in the high dose administration group (P<0.05). Pathological sections of liver tissue showed that compared with the model group, the vacuolar edema, inflammatory cell infiltration and lipid droplets of hepatocytes in the administration group were significantly reduced. Through network pharmacology, 13 key active components such as isoprenin and acacetin and 50 key targets such as AKT1, GAPDH, TNF were screened from the total flavonoids of Chuju. Ultimately, it was postulated that the total flavonoids of Chuju might improve hypercholesterolemia through the PI3K-AKT signaling pathway and endocrine resistance pathway. Conclusion: Total flavonoids of Chuju improved hypercholesterolemia induced by a high-fat diet in mice. The mechanism of action appeared to involve 13 key components acting on 50 targets, which played an antioxidant role and reduced lipid peroxide production through key pathways such as the PI3K-AKT signaling pathway. The final result was prevention of excessive cholesterol accumulation in vivo.