Background Metabolic comorbidities associated with heart failure with preserved ejection fraction (HFpEF) have complicated the treatment of this debilitating syndrome. Systemic metabolic dysregulation, cardiac dysfunction, and elevated oxidative stress in cardiometabolic HFpEF manifest in severe exercise intolerance, diminished quality of life, and repeated hospitalizations. Renal denervation (RDN) is a device-based therapy designed for treatment of resistant hypertension. RDN has recently been shown to attenuate the severity of metabolic syndrome in preclinical models. To date, RDN has not been evaluated in the setting of severe HFpEF. Objective To investigate the effects of RDN on exercise tolerance, vascular reactivity, and skeletal muscle mitochondrial function in a clinically relevant cardiometabolic model of HFpEF. Methods 20-week-old male ZSF1 obese rats (n=13-14 per group) were randomized to undergo either Sham-RDN or radiofrequency (RF)-RDN treatment on week 4 of a 10-week HFpEF protocol. At baseline, 4-, and 8-weeks of the study protocol, rats were subject to a treadmill exercise protocol. At 10 weeks, thoracic aorta and gastrocnemius muscle samples were collected for vascular reactivity and mitochondrial respiration studies, respectively. Results Compared to the sham-RDN group, RDN-treated ZSF1 obese rats demonstrated significant improvement in exercise capacity at 4 weeks following treatment (Figure A). Acetylcholine-induced vasorelaxation was significantly (p < 0.01 vs. Sham-RDN) following RDN treatment (Figure B). Furthermore, we observed significantly (p < 0.01) improved respiration (State 3) in skeletal muscle mitochondria isolated from RDN-treated animals compared to sham-RDN (Figure C). Conclusion Our data indicate that RDN improves exercise tolerance in the setting of severe HFpEF. Ablation of the renal sympathetic nerves enhanced vascular function which is critical for exercise performance. The improvement in skeletal muscle mitochondrial respiration that we observed likely contributes to the improved exercise tolerance following RDN. These provocative findings suggest that inhibition of renal sympathetic nerve activity by RDN improves peripheral vascular and skeletal muscle function to improve exercise capacity. Furthermore, these findings strongly support RDN as a viable therapy to treat patients that suffer from cardiometabolic HFpEF.