The inhibitory effects of 1,3-diacylglycerol (DAG) on diet-induced lipid accumulation in liver and abdominal adipose tissue of rats were investigated in the present study. Male Sprague-Dawley rats were given free access to diets containing 7 wt% TAG (low TAG), 20 wt% TAG (high TAG), or 20 wt% DAG (high DAG), respectively, for 8 wk. The body weight of rats in the 20% high-TAG group increased significantly, and the weights of their abdominal adipose tissue and liver also showed a significant increase compared with rats in the low-TAG group. However, the high-DAG diet resulted in both a significant reduction in body weight gain (with a decrease of 70.5%) and an increase in the ratio of abdominal fat to body weight (by 127%) compared with the high-TAG diet. As well, the liver TAG and serum TAG levels of the high-DAG group were significantly lower than those of the high-TAG group. These effects were associated with up-regulation of acyl-CoA carnitine acyltransferase (ACAT) and down-regulation of acyl-CoA DAG acyltransferase (DGAT) in the liver. However, no significant difference was observed in the activities of alanine aminotransferase and aspartate aminotransferase among the groups (P > 0.05). The present results indicate that dietary DAG reduced fat accumulation in viscera and body, and these effects may be involved with up-regulation of ACAT and down-regulation of DGAT in the liver.