Objective To investigate the effect of TMAO, a metabolite of intestinal flora, on hepatic lipid metabolism in rats with splenic deficiency and hyperlipidemia, and to further explore the possible mechanism of Xiangsha Liujunzi Decoction in the intervention of hepatic lipid metabolism disorder. Methods SD rats were divided into: (1) Blank control group (group C), (2) blank control group DMB (group C D), DMB is TMAO inhibitor, (3) spleen deficiency hyperlipidemia group (PG group), (4) spleen deficiency hyperlipidemia DMB group (PG D), (5) Spleen deficiency hyperlipidemia Xiangsha Liujunzi Decoction group (PG XS group), Except group C and group C D, the other groups were constructed to establish spleen deficiency hyperlipidemia model (12 weeks of modeling) by combining excessive fatigue and high-fat diet. After model establishment, group C D and PG D were given 1%DMB in drinking water every day, and group PG XS was given Xiangsha Liujunzi decoction (11.34 g crude drug /kg/ day) every day. The other groups were given the same amount of normal saline. The blood lipid level of each group was detected by automatic biochemical method after 4 weeks of intragastric administration. The morphological changes of liver were observed by HE staining. The lipid deposition in the liver of each group was observed by oil red O method. Liver FFA, TG and TC were detected by ELISA. The plasma TMAO content was detected by LC-MS. The relative expression levels of PERK, FOXO1, SREBP-2, ABCA1 and miR-33 mRNA in liver were detected by qRT-PCR. The contents of SREBP-2 and ABCA1 gene protein in liver were detected by WB. Results (1) The content of TC, TG and LDL-C in serum of PG group was significantly higher than that in C group, and the content of HDL-C was significantly lower than that in C group; The contents of FFA, TG and TC in liver tissue of PG rats were significantly increased compared with those of C group. Compared with group C, lipid deposition in liver was aggravated and fat vacuoles were increased significantly. There was no difference between group C D and group C in the above indexes. Compared with the PG group, PG D and PG XS groups could significantly reduce the contents of TC, TG and LDL-C in serum, increase the content of HDL-C, reduce the contents of FFA, TG and TC in liver tissue, alleviate the phenomenon of lipid deposition in liver tissue and reduce liver vacuoles. There was no significant difference between PG D group and PG XS group. (2) Compared with group C, the plasma TMAO content of PG group was significantly increased, the liver PERK, FOXO1 and miR-33a mRNA expressions were significantly increased, and the liver SREBP-2 and ABCA1 mRNA and protein expressions were significantly decreased; There was no difference between group C D and group C in the above indexes. Compared with the PG group, PG D and PG XS groups could significantly reduce the plasma TMAO content, decrease the liver PERK, FOXO1 and miR-33a mRNA expressions, and increase the liver SREBP-2 and ABCA1 mRNA and protein expressions. There was no significant difference between PG D group and PG XS group. Conclusion TMAO may regulate the SREBP-2/miR-33a/ABCA1 signaling pathway through PERK/FOXO1 axis to cause liver lipid metabolism disorders in rats, and Xiangsha Liujunzi Decoction may inhibit liver lipid metabolism disorders by inhibiting TMAO content.