To investigate the molecular mechanism by which diets of different energy levels alter fat accumulation, 120 Black-Bone chickens were divided into 3 groups and fed diets with varying energy levels for six weeks (low: 3.02 Mcal/kg, moderate: 3.22 Mcal/kg, high: 3.42 Mcal/kg). The high-energy diet increased the concentration of blood lipids. Furthermore, high-energy intake inhibited the activities of several serum lipases. Histological sections showed over-synthesis of lipids in the livers of chickens fed high-energy diets. Data from western blotting and PCR analyses indicated that key factors for lipogenesis, regulatory transcription factors and fatty acid transporters, were up-regulated, while key factors for lipolysis were down-regulated. Chickens fed low-energy diets showed opposite results. In conclusion, varying energy levels of diets affect fat accumulation in Black-Bone chickens through changed expression of metabolic regulators.