Introduction Clinical use of Doxorubicin (DOX), a widely useful drug for the treatment of various cancers, is limited by the cardiotoxicity. Despite the growing attention to the protective role of exercise activity, especially moderate endurance exercises against the DOX-induced cardiotoxicity, few studies have focused on aerobic high-interval interval training (AIT) and its mechanisms. Therefore, the purpose of this study was to investigate the protective effect of AIT against the DOX-induced changes in the PGC-1α mRNA levels in the cardiomyocytes of rats. Methods 24 male Wistar rats were randomly assigned into four groups (n=6/groups): 1) Control; 2) DOX (20 mg/kg body weight)/; 3) AIT (7 sets of 4 min intervals at 80%–90% VO2max interspersed with 3 min periods of 65%–75% VO2max, for 8 weeks before DOX-injection); and 4) AIT + DOX. The mRNA levels also were determined using RT-PCR. For statistical analysis of data, One-way analysis of variance and Tukey's post hoc test were used (α<0.05). Results DOX-treatment significantly decreased the expression of PGC-1α in the DOX group compared to the Control group (P < 0.05). Also, the expression of PGC-1α significantly increased in the AIT group compared to the Control group (P < 0.05). AIT before DOX-induction also significantly increased the expression of PGC-1α in the AIT+DOX group compared to the DOX group (P < 0.05). AIT could inhibit the DOX-induced changes in the PGC-1α mRNA levels in the cardiomyocytes of rats. Conclusion AIT could be a good non-prescriptive strategy for preventing of DOX-induced cardiotoxicity.