STUDY OBJECTIVES: Restless legs syndrome (RLS) is known to be a risk factor for cardiovascular disease. However, there are no electrophysiological biomarkers to assess this risk. This study aimed to evaluate heart rate variability (HRV) and cardiovascular reflexes in the supine and standing positions during wakefulness in patients with RLS. METHODS: Fourteen drug-naive patients with RLS (12 women and 2 men, mean age, 42.14 +/- 7.81 years) and 10 healthy control patients underwent tests for blood pressure, heart rate when in the supine and standing positions, and deep breathing and handgrip tests in controlled laboratory conditions. Data on 5-minute R-R intervals at each position were collected and analyzed for HRV. RESULTS: Expected cardiovascular reflexes were within the normal range and were similar between the 2 groups. In HRV analysis, the normalized unit of the low-frequency component and the low-frequency/high-frequency ratio during standing were lower in patients with RLS than in the control patients. The low-frequency/high-frequency ratio responses during the change from the supine to the standing position were significantly reduced in patients with RLS (mean +/- standard deviation, 2.94 +/- 3.11; control patients: 7.51 +/- 5.58; P = .042.) On Spearman rank correlation, questionnaires related to sleep problems were associated with the parameters of HRV. CONCLUSIONS: Patients with RLS showed reduced sympatho-vagal responses during the change from the supine to the upright position during wakefulness, and RLS-related sleep disturbance was a contributing factor for autonomic nervous system dysfunction. This case-control study showed a difference in HRV response to position change in a considerably small group of patients with RLS. The relevance of this finding is uncertain, but it may be worthy of further investigation in longitudinal studies on RLS and cardiovascular disease.