The present study compared the effects of dietary diacylglycerol (DG) and triacylglycerol (TG) oil on lipid metabolism in rats fed a 5% fat (AIN-76) diet for 6 weeks. The plasma and hepatic lipids, hepatic cholesterol-regulating enzyme activity, and hepatic and adipose tissue fatty acid metabolism enzyme activities were determined. Among plasma lipids, triglyceride, free fatty acid, and phospholipid concentrations were significantly lower in the DG group than in the TG group. A lower plasma TG level was accompanied by an increase in adipocyte lipoprotein lipase activity. The hepatic triglyceride level was significantly (P < .001) lowered in the DG group, which was attributable to an increased fatty acid oxidation enzyme (beta-oxidation) activity and a reduced fatty acid synthesis enzyme (glucose-6-phosphate dehydrogenase) activity. The plasma cholesterol concentration was significantly lower in the DG group and was accompanied by a lower hepatic 3-hydroxy-3-methylglutaryl-coenzyme A reductase activity. The DG oil used in this study was beneficial for enhancing lipid metabolism with apparent hypolipidemic effects.