Remote ischemic preconditioning (RIPC), elicited by brief, intermittent periods of ischemia followed by reperfusion, has cardioprotective effects. Our lab recently found that repeated bouts of RIPC improves microvascular function; however, it is unclear whether autonomic function would also be affected. We hypothesize that heart rate variability (HRV) will be altered following two weeks of repeated RIPC. Eleven healthy adults (age = 22 ± 2 years, 6 males, 5 females, BMI = 23.1 ± 3.4) performed two weeks of RIPC over 3 four-day periods, each separated by a one-day break. Each RIPC session consisted of 4 repetitions of 5 minutes of arm blood flow occlusion interspersed by 5 minutes of reperfusion. Before and one day after two weeks of RIPC, resting heart rate was collected with lead II electrocardiogram (ECG) while the participant was comfortably lying in a semi-recumbent position in a quiet, dimly lit thermoneutral environment. Power spectral density and symbolic dynamics were used to assess HRV from ~350 beat ECG segments. Two weeks of repeated RIPC decreased the percentage of 0V fragments (pre-RIPC = 13.1 ± 6.5%, post-RIPC = 6.9 ± 4.9%, P < 0.05) and increased the percentage of 2V fragments (pre-RIPC = 42.9 ± 12.1%, post-RIPC = 52.5 ± 10.0%, P < 0.01), while decreasing the LF/HF ratio (pre-RIPC = 1.1 ± 0.6, post-RIPC = 0.7 ± 0.3, P < 0.01). Thus, both the spectral and fractal analyses indicate that repeated RIPC shifts sympathovagal balance by increasing parasympathetic and decreasing sympathetic activity.