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Background & Aims: The expression of myocardium gene can be affected by endurance activity; besides, sox6 transcription factor influences its formation. This study aimed to investigate the effect of 14 weeks of endurance activity on expression of sox6 gene of left ventricle in male Wistar rats. Methods: Forteen rats were housed under the controlled conditions and after adjusting with endurance program, were randomly assigned into two groups of control and experimental. For the experimental group, an endurance program (14 weeks, 6 days/week, 60 minutes/day, and velocity of 30 m/minutes) on motorized treadmill was performed. Then, the rats were anesthetized and sacrificed 48 hours after the end of the last session and the left ventricle was removed. Real-time polymerase chain reaction (RT-PCR) method was used to determine the expression levels of sox6 gene in the left ventricle. The collected data were evaluated using t-test. Results: The ratio of left ventricle weight (LVW) to the body surface area (BSA) was significantly (P=0.010) higher in experimental group (0.168 ± 0.008 g/m2 ) compared to the control group (0.153±0.006g/m2 ). The ratio of heart weight (HW) to the body surface area was significantly (P = 0.002) higher in experimental group (0.270 ± 0.014) compared to the control group (0.240 ± 0.019), too. In addition, the mean expression value of sox6 gene of left ventricle was significantly higher in experimental group (P = 0.001), too. Conclusion: It seems that physical activity improves heart functional indices (the ratio of left ventricle and heart weight to body surface area), especial in left ventricle, via increasing the expression ofsox6 gene.