Objective To explore the safety and efficacy of the renal sympathetic denervation under the guidance of EnSite-NavX three-dimensional mapping system. Methods A three-dimensional model of abdominal aorta and renal artery was constructed under the guidance of EnSite-NavX three-dimensional mapping in 6 healthy Guangxi Bama mini-pigs. Then a radiofrequency ablation catheter was put into the bilateral main trunks of renal artery and six points were marked within each trunk from far to near the heart. One side of the renal artery was ablated and the other received no ablation serving as control. The mini-pigs were followed up for 6 months after the procedure. The norepinephrine (NE) content in the renal tissues was detected by ELISA. Sympathetic nerve damage in the renal tissue of the ablation side was observed by Haematoxylin-Eosin staining. Results Modeling and unilateral renal sympathetic denervation were successfully performed in the six mini-pigs. Subcutaneous hematoma was formed after operation in one pig. None of the pigs had the complications such as renal artery stenosis, thrombopoiesis, or dissection. The NE content of ablation side was significantly lower than that of the opposite side ([170.13±52.60] ng/g vs [429.86±131.43] ng/g, P<0.05). Haematoxylin-Eosin staining showed broken epineurium and loss of axons in the renal sympathetic nerve of the ablation side. Conclusion A three-dimensional model of abdominal aorta and renal artery can be clearly constructed under the guidance of EnSite-NavX three-dimensional mapping. Renal sympathetic denervation can be achieved safely and effectively without fluoroscopy or renal arteriography under the navigation of NavX.