BACKGROUND: The expression and activity of fatty acid synthase (FAS) enzymes determine de novo fatty acid synthesis, which can be enhanced by a high-fat and high fructose diet or inhibited by some bioactive compound diets. Beets are a great source of therapeutic compounds that have the potential to improve health and prevent disease. AIM: This study examined the effects of beets on liver FAS gene expression and FAS levels. METHODS: A total of 25 male Wistar rats divided into five groups: (1) Standard diet (n); (2) high fat and fructose diet (HFFD); (3) HFFD have given beet 6%-contained standard diet (B1); (4) HFFD have given beet 9%-contained standard diet (B2), and (5) HFFD have given beet 12%-contained standard diet (B3). The HFFD was given to rats in the 2, 3, 4, and 5 group diets for 8 weeks? and then 3, 4, and 5 groups received beet-contained standard diet for 6 weeks. At the end of the intervention, FAS levels were measured (please specify where it was measured from) using the ELISA method, liver FAS gene expression was analyzed by quantitative polymerase chain reaction, and triglyceride (TG) levels were determined by the colorimetric method. RESULTS: The beet-substituted diet significantly suppressed the hepatic FAS gene expression and decreased the serum FAS levels in rats previously given HFFD (p < 0.05). The expression of the FAS gene showed a significant positive correlation with the levels of FAS serum (p < 0.05), and also with the hepatic TG levels but not significant (p > 0.05). Substitution of beet 9% in diet gives the best effect in hepatic FAS gene expression and the serum FAS levels. CONCLUSIONS: The diet contained beet 9% was seen as a necessary physiological dose to improve the effects of high-fat and diet fructose diet through suppressing FAS gene expression and a decreased serum FAS levels.