Effect of chicory root extract (CRE) on the triglyceride metabolism in orotic acid (OA)-fed rats was investigated. Liver weights and hepatic triglyceride concentrations were markedly increased by OA-feeding rats. These results were attributed to the significant increase in the activity of hepatic phosphatidate phosphohydrolase (PAP), diacylglycerol acyltransferase (DGAT), and ratelimiting enzymes for triglyceride synthesis. Supplementation of CRE to OA did significantly reduced the hepatic triglyceride concentrations and DGAT activity without affecting PAP activity. Furthermore, OA treatment was significantly decreased plasma triglyceride (TG) and increased hepatic TG concentrations and reduced microsomal triglyceride transfer protein (MTP) activity without diminishing MTP mRNA expression in rats. However, hepatic TG concentration was significantly decreased and MTP activity was also reduced without diminishing MTP mRNA expression in rats fed simultaneous with OA and CRE diet. The hepatocytes in the OA-feeding rats contained numerous largely fat droplets, but CRE feeding prevented the OA-induced fat accumulation. Present study demonstrates that CRE reduces the liver TG accumulation by reduced DGAT and MTP activities without diminishing MTP mRNA expression by OA administration.