The purpose of this study was the effects of exercise preconditioning with high intensity interval training (HIIT) on metabolites of NO (NO2-, NO3-), and infarction size of myocardium after ischemia/reperfusion injury in healthy male rat. For this reason, 44 wistar rat were randomly divided into 4 group HIIT (n=8), Group HIIT + protocol IR (n=14), control group (n=8) and control + protocol IR (n=14). Each session of HIIT was 1 hour of exercise in the 3-stage, 6-minute running of 50 to 60% VO2max warm up, 7 Period 7-minute running with an including of 5 to 20 ° slop (4 min with an intensity of 80 to 120, 3 minutes of 50-60% VO2max) and 5-minute running of 50 to 60% VO2max cool-down. Control group did not participate in any exercise program. NO and its metabolites were measured by using Griess reaction. Results showed there was a significant difference on nitrite (0. 855 mol per liter, equivalent to 34. 79%), nitrate (6. 202 mol per liter, equivalent to 149. 48%) and NOX (6. 6 micromoles per liter, equivalent to 98. 11%) levels in HIIT group than control group. That, the results showed, AAR was significantly more (9. 06%, p=0. 010) and INF was significantly lesser (23. 2%, p <0. 001) in exercise training group in comparison with control. In generally, it seems that increase in No-NO3-, NO2-axis is one of mechanisms that HIIT exercise program can protect heart from Ischemia/Reperfusion injury and decreased myocardial infarction.