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Abstract. lifestyle-related diseases are associated with overeating and lack of exercise. the purpose of this study was to investigate the effect of exercise and high-fat diet on plasma adiponectin and nesfatin levels. mice were housed for 4 weeks in 4 groups, which included the non-exercise and normal diet (Sn), exercise and normal diet (En), non-exercise and high-fat diet (SF) and the exercise and high-fat diet (EF) group. the mice in the exercise groups were housed in cages with a running wheel and were subjected to voluntary exercise. the food intake (Kcal) of the mice in the exercise groups increased compared to that of the mice in the non-exercise groups (p<0.01). Body weight and visceral fat decreased in the mice in the EF group compared to the mice in the SF group (p<0.01 and p<0.05). the temperature of the mice in the EF group increased compared to that of the mice in the Sn group (p<0.05). Blood glucose, insulin (p<0.01), cholesterol (p<0.01) and triglyceride concentrations (p<0.01) increased in the SF group compared to the normal diet groups. Furthermore, plasma insulin and cholesterol concentrations increased in the SF group compared to the exercise groups (p<0.01). plasma adiponectin and nesfatin-1 levels in the SF group decreased compared to the Sn group (p<0.05). Exercise under a high-fat diet antagonized the significant decrease in the nesfatin-1 level. Exercise together with a high-fat diet affected the plasma levels of adiponectin and nesfatin. it is therefore suggested that exercise together with a high-fat diet can affect various diseases via adiponectin and nesfatin.