Purpose: Although physical activity is one of the medical aid strategies to reduce arterial stiffness in diabetic patients, the mechanism remains unclear. The purpose of this study was to determine the effect of aerobic exercise on Klotho-fibroblast growth factor-23 (FGF23) axis and arterial calcification in type 2 diabetic postmenopausal women. Methods: Twenty-four type 2 diabetic postmenopausal women who were registered at the diabetes association of Shahrood, were randomly assigned into two groups including exercise (n=12, 51± 6 yrs., 31. 9± 5 kg/m2) and control (n=12, 52± 5 yrs., 29. 23. 8 kg/m2). The aerobic training consisted of 45 to 60 minutes running with 45 to 70 percent of heart rate reserve, three sessions per week for 8 weeks. Before and after the experimental period, blood fasting samples were taken and the ELISA method was used to analyzed Klotho-FGF23. Arterial stiffness analysis was also performed using VaSera-VS-2000. The data were analyzed using the statistical tests of Paired and Independent t-test (P<0. 05). Results: No significant changes in phosphor, FGF23, phosphor × calcium, diastolic blood pressure and VO2max were found in this study (P>0. 05). But, calcium levels in exercise group decreased significantly following the experimental period (P<0. 05). Moreover, systolic blood pressure, Klotho-FGF23 and arterial stiffness significantly decreased and klotho significantly increased in exercise group (P<0. 05). Conclusion: Aerobic training can decrease the arterial stiffness by modifying the underlying causes of SBP, Klotho and Klotho-FGF23 in postmenopausal women with diabetes. However, due to the few studies in this regard, these findings need to be validated in the future studies.