This study purposed to examine the effect of puer-tea supplementation on antioxidant enzymes during acute exercise of rats. Thirty-two rats were randomly divided into four groups (HS : high fat diet sedentary group, n=8; HE : high fat diet acute exercise group, n=8; PS : puer-tea gavaged sedentary group, n=8; puer-tea gavaged acute exercise group, n=8), The high-fat diet was composed of 35% carbohydrate, 20% protein, and 35% fat. In the acute exercise group, the rats were swam for 60min (3% load/weight) before anesthetized with ether. The results showed that EFT of PS group was significanlty lower than the HS group (p < .05). In the blood components, there were no significantly different in the serum glucose, TC, HDLC, but TG level of exercise group were significantly lower than the control group (p < .01), respectively. In addition muscle glycogen content of PS and PE were significantly higher than the HS and HE (p < .01), respectively, and MDA content of PE was lower than the PS group (p < .01). Furthermore, Cu,Zn-SOD protein expression of PS group was significantly higher than the HS group (p < .05), and GPx protein expression of PE and HE were significantly higher than the PS and HS (p < .01), respectively. From these results, puer-tea supplementation has a positive effect on antioxidant activity because of puer-tea supplemented group was showed higher value of Cu,Zn-SOD protein expression, and could be an important dietary antioxidant source as well.