Medium chain triglyceride (MCT) is dietary fat naturally existed in palm oil and breast milk. As a novel functional ingredient, it can be used in the production of formula food for special medical purpose. However, the specific target for MCT regulating lipid metabolism is still unclear. Based on network pharmacology analysis and rat model validation, the potential role and target of MCT for regulating lipid metabolism were explored. Animal model demonstrated that no hepatorenal toxicity in rat was observed after nutritional intervention of MCT, indicating its good edible safety. And MCT nutritional intervention down-regulated the levels of serum triglyceride, total cholesterol and low-density lipoprotein, and inhibited the accumulation of adipose tissue of the epididymis in rat. Network pharmacology analysis showed that there were 27 overlapping targets between MCT metabolites (octanoic acid and decanoic acid) and lipid metabolism-related genes, and potentially involved in regulation of peroxisome proliferator activated receptor lipid metabolism-related pathways and fatty acid binding and transport. Western blot assay also indicated that MCT could down-regulate the expression of fatty acid-binding protein 4 (FABP4) of the small intestine in a dose-dependent manner. Results indicated that the MCT nutritional intervention could down-regulate the expression of FABP4, subsequently affecting the absorption and transport of fatty acid, reducing the level of serum lipids and alleviating fat accumulation, thereby regulating the lipid metabolism. The purpose of this study was to clarify the role of MCT in regulating lipid metabolism and its potential targets, and to provide a theoretical basis for the application of MCT in lipid metabolism regulation.