To study effect of neuronavigation-assisted microscopic transsphenoidal resection on autonomic function related hormone levels and visual function in patients with pituitary adenoma. Methods: A total of 87 patients with pituitary adenoma were divided into the observation group (45 cases) and control group (42 cases). The control group underwent neuroendoscope-assisted endoscopic transsphenoidal resection, and the observation group underwent neuronavigation-assisted microscopic transsphenoidal resection. Patients were followed up for 1 month, and surgery-related indexes, pituitary function-related hormone levels, visual function, and complications were compared between the two groups. Results: The observation group's operation time and hospital stay were significantly shorter and blood loss was significantly greater than those of the control group (P<0.05, P<0.01). Thyroid stimulating hormone (TSH), free T3 (FT3), adrenocorticotropic hormone (ACTH), and prolactin (PRL) levels were significantly higher and cortisol (COR) level was significantly lower in the observation group than in the control group (all P<0.05). Mean defect (MD) and pattern standard deviation (PSD) of the visual field were significantly lower and visual field index (VFI) was significantly higher in the observation group than in the control (P<0.05, P<0.01). There was no significant difference between the two groups in cerebrospinal fluid leakage and complications (P>0.05). Conclusion：Neuronavigation-assisted microscopic transsphenoidal resection helps to promote the recovery of pituitary function-related hormone levels and improve visual function.