Perilla frutescens seeds are rich in proteins and their amino acid composition is comprehensive， so， the active peptides of Perilla frutescens seeds have great development and utilization value. In this work， the Perilla frutescens seed peptides were prepared by enzymatic hydrolysis and the obtained peptides was set as low， middle and high doses. After oral administration of Perilla frutescens seed peptides for 28 days， the weight loading swimming time of male mice was investigated， and muscle/liver glycogen， blood lactic acid （BLA）， blood urea nitrogen （BUN）， and lactate dehydrogenase （LDH） were further analyzed. The results show that compared with the control group， the swimming time in low， medium and high dose groups increased by 10.61%， 22.76% and 56.14%， respectively. Perilla frutescens seed peptides significantly （P<0.05） increased the levels of muscle/liver glycogen accumulation， improved serum LDH vitality， and accelerated the catabolism of BLA and BUN. Therefore， Perilla frutescens seed peptides exhibited excellent anti-fatigue activity.