Objective To explore the effect of evidence-based protocol on reducing medical adhesive-related skin injury (MARSI) in neonatal intensive care unit (NICU). Methods A total of 308 newborns who used medical adhesives were chronologically divided into a control group and an observation group, with 154 in each group. The control group was given conventional nursing care, while the observation group received evidence-based nursing which was guided by a protocol regarding MARSI prevention in NICU. The incidence of MARSI and pain intensity after removal of the adhesives were compared between the two groups. Cognitive and behavioral changes of medical staff before and after the evidence-based practice were also evaluated. Results The observation group had lower incidence of MARSI and milder pain intensity compared with the control group (P<0.01 for both). After implementation of the evidence-based practice, MARSI knowledge and behaviors of pediatricians and nurses were significantly improved (P<0.05, P<0.01). Conclusion Implementation of evidence-based protocol can reduce the incidence of MARSI in NICU.