Background and Aims: Heart rate variability (HRV), i.e., the beat-by-beat fluctuations in heart rate (HR) reflecting the autonomic nervous system balance, is altered in patients with diabetes. This has been associated with arterial aging (stiffer arteries) and differs in men and women. The present study hypothesized that the impact of HRV on arterial aging, indexed as carotid-femoral pulse wave velocity (PWV), differs in a gender-specific manner and is affected by diabetes mellitus. Method: A total of 422 outpatients (187 women and 235 men) were studied. PWV was measured using the validated SphygmoCor device (AtCor Medical). Time-domain and frequency-domain parameters were measured to assess HRV. Results: The prevalence of diabetes was 30.8% with a slight, but nonsignificant, greater prevalence in men. Both age and SBP were independent determinants of PWV in each of the four groups (men and women with or without diabetes). Low-frequency activity was inversely correlated with PWV. It was greater in women without diabetes, but it was not significant in men regardless of the presence of diabetes. Conclusions: Beyond age, blood pressure, and diabetes, impaired cardiac autonomic function assessed by determination of HRV was significantly associated with arterial aging. The association between lower sympathetic and parasympathetic activity and stiffer arteries was significant in women, but not in men.