Stigmasterol (ST) improves lipid and bile acid (BA) metabolism. This study investigated ST's mechanism in preventing dyslipidemia via BA metabolism. ST treatment alleviated high-fat diet (HFD)-induced lipid metabolism disorder. Fecal microbiota transplantation from ST-treated rats showed similar protective effects. ST reversed HFD-induced gut microbiota dysbiosis, modifying serum and fecal BA metabolome profiles, particularly CYP7A1-mediated pathways. Chenodeoxycholic acid (CDCA) combined with ST amplified therapeutic effects. ST benefits against HFD-induced hyperlipidemia and obesity are partially due to intestinal microbiota reprogramming and BA metabolism in enterohepatic circulation. This suggests the gut microbiota as a potential target for HFD-induced dyslipidemia treatment.