Dingxin recipe Ⅲ ameliorates atherosclerosis through stard4-mediated regulation of hepatic lipid metabolism

BACKGROUND Atherosclerosis is the pathological basis of cardiovascular diseases. Dingxin Recipe III (DXRIII), a traditional Chinese herbal formula, has shown therapeutic effect for atherosclerosis, though its mechanisms remain unclear. This study aimed to investigate the effects and molecular mechanisms of DXRIII on atherosclerosis progression. METHODS Male ApoE-/- mice were fed a high-fat diet for 12 weeks to induce atherosclerosis, followed by 12 weeks of treatment with DXRIII (7.5 or 15 g/kg/d), atorvastatin, or saline. Serum lipids, liver enzymes, aortic plaques, and hepatic lipid deposition were assessed. Transcriptomics, proteomics, and metabolomics analyses identified hepatic molecular changes. Key targets were validated by western blot, RT-qPCR, immunohistochemistry, and hepatocyte models. Molecular docking and cellular thermal shift assay assessed the direct binding of DXRIII components to target proteins. Gene overexpression and knockdown experiments were conducted in vitro and in vivo. RESULTS DXRIII significantly reduced aortic plaque areas, improved lipid profiles (decreased triglycerides, total cholesterol, and low-density lipoprotein-C), and alleviated hepatic steatosis. Integrated multi-omics revealed modulation of lipid metabolism pathways, including steroid hormone biosynthesis and arachidonic acid metabolism pathways. Steroidogenic acute regulatory-related lipid transfer protein 4 (Stard4) was identified as a key target, with expression positively correlated with gamma-linolenic acid and negatively correlated with corticosterone. Direct binding between DXRIII components and Stard4 was observed. Stard4 overexpression reduced lipid accumulation, while knockdown aggravated lipid deposition and negated the effect of DXRIII. Hepatic Stard4 knockdown aggravated atherosclerosis and lipid-related genes expression (Angptl4, Apob, Soat2, Scarb1, Lepr). CONCLUSION DXRIII attenuates atherosclerosis by upregulating hepatic Stard4 expression to restore lipid homeostasis and reduce lipid accumulation.