The present work aimed to investigate the impact of intake of diacylglycerol(DAG) from soybean oil on the reduction of fat accumulation and the long-term effects of dietary intake of DAG and triacylglycerol(TAG) with similar fatty acid compositions on the development of obesity. Kuming mice were used to compare the effects of low-dose TAG(2.5 g/kg BW), low-dose DAG(2.5 g/kg BW), high-dose TAG(10 g/kg BW), and high-dose DAG(10 g/kg BW) on the induced obesity. The results showed that the body weight and serum triglyceride concentration decreased significantly in both DAG-treated groups compared with the TAG-treated groups(P < 0.01, P < 0.05, respectively). However, the plasma glucose concentration was significantly lower in the DAG-treated groups than the TAG-treated groups(P < 0.05); the weight and morphology of the liver and kidney in DAG-treated groups were similar to those in the control group, there were no significant differences within each group. The present results indicated the anti-obesity and lipid-lowering effects of dietary DAG oil in mice and its potential use as a functional food for humans.