Obesity has been defined as a state of continuous oxidative stress. There are numerous approaches wherein weight problems immediately affect the cardiovascular device. The klotho gene acts as aging -suppressor gene that protects cells and tissues from oxidative stress. It’s now widely known that exercise or antioxidants can prevent oxidative harm by decreasing oxidative stress. This study intended to clarify the effects of exercise and antioxidants on klotho gene mRNA expression in rat’s cardiac tissue during oxidative stress .The animals have been divided into five groups. The group I serve as negative control. Group II is a positive control feeding high fat diet (HFD) (Oxidative stress group) .Group III: (HFD) treated with exercise training. Group IV: (HFD) treated with vitamin E as antioxidant and group V: (HFD) treated with exercise training and vitamin E. Serum insulin, glucose, HOMA, total cholesterol and TG were measured. Gene expression of TNFα mRNA expression level in cardiac tissue and Klotho mRNA expression in kidney tissue were performed using Real-time PCR. We demonstrate that combined exercise training with vitamin E treatment for 8 weeks resulted in a significant improvement in the insulin resistance and dyslipidemia. Also, it was associated with significant decrease in TNFα expression in heart and increase in Klotho gene expression in kidney. We concluded that the exercise training and /or antioxidant treatment improved oxidative stress effects in cardiac tissue through up-regulation of mRNA expression of Klotho gene in the kidney and down-regulation of TNFα expression in the heart.