This study investigates the mechanism of Xiangsha Liujunzi Decoction in regulating HDL metabolism disorders in the liver of rats with impaired lipid transportation due to spleen dysfunction, by targeting the TMAO-mediated PERK/FOXO1 axis. Thirty-five SD rats were divided into five groups: a normal group (group Z), a TMAO inhibitor DMB group (group D), a spleen dysfunction with impaired lipid transportation model group (group PSJY), a model combined with DMB intervention group (group PSJY+D), and a model combined with Xiangsha Liujunzi Decoction intervention group (group PSJY+XS). Except for groups Z and D, all other groups were subjected to over-fatigue combined with a high-fat diet for 12 weeks to establish the model. Starting from week 13, 1% DMB was added to the drinking water for groups D and PSJY+D, while group PSJY+XS was administered Xiangsha Liujunzi Decoction (11.34 g crude drug/kg/day) by gavage for 4 weeks. The experimental results show that compared with group Z, group PSJY exhibits significant hepatic lipid deposition. Serum levels of TC, TG, LDL-C, and ApoB are significantly increased (P < 0.01), while HDL-C and ApoA-I are significantly decreased (P < 0.01). Plasma TMAO levels are elevated (P < 0.01). The protein expressions of p-PERK and FOXO1 in liver tissue are increased (P < 0.01), whereas the mRNA and protein expressions of SR-BI, ApoA-I, and HL are decreased (P < 0.01). In the DMB intervention group, except for reduced TMAO and downregulated SR-BI mRNA expression, no significant changes are observed in other indicators.Compared with group PSJY, group PSJY+D shows reduced hepatic lipid droplets. Serum TG and ApoB are decreased (P < 0.01), while ApoA-I is increased (P < 0.05). TMAO is significantly reduced (P < 0.01). The protein expressions of SR-BI, ApoA-I, and HL are increased (P < 0.05 or P < 0.01). Group PSJY+XS also shows alleviated hepatic lipid deposition. Serum TG, LDL-C, and ApoB are decreased (P < 0.01), while HDL-C and ApoA-I are increased (P < 0.01, P < 0.05). TMAO levels are reduced (P < 0.01). The protein expressions of p-PERK/PERK and FOXO1 are decreased (P < 0.05). The mRNA and protein expressions of SR-BI, ApoA-I, and HL are significantly increased (P < 0.05 or P < 0.01). In conclusion, under the condition of impaired lipid transportation due to spleen dysfunction, changes in the TMAO/PERK/FOXO1 axis can lead to downstream HDL metabolic disorders. Xiangsha Liujunzi Decoction can alleviate HDL metabolism disorders in this model by targeting the TMAO-mediated PERK/FOXO1 axis.