To assess ICU nurses′ knowledge, attitude, and practice (KAP) regarding non-pharmacological interventions for acute gastrointestinal injury (AGI), and to analyze the influencing factors,thereby providing a reference for developing systematic training programs. Methods A total of 237 ICU nurses from 16 general hospitals in Shenzhen were conveniently selected as participants, then they were investigated by using a self-administered questionnaire of ICU nurses′ KAP regarding non-pharmacological interventions for AGI, which was developed based on AGI expert consensus and the KAP framework. Results In the KAP questionnaire for ICU nurses regarding non-pharmacological interventions for AGI, the participants′ knowledge dimension was 16.0 (8.0,19.0) points, attitude dimension was 36.0 (30.0,39.0) points, and practice dimension was 36.0 (32.0,42.0) points. Multiple linear regression analysis showed that, knowledge was influenced by education level, critical care specialist nurse or not, AGI patients care experience, satisfaction with professional growth in ICU, and satisfaction with personal competency utilization;attitude was influenced by critical care specialist nurse or not, AGI patients care experience, satisfaction with professional growth in ICU and satisfaction with personal competency utilization; practice was influenced by satisfaction with professional growth in ICU and satisfaction with personal competency utilization (all P<0.05). Conclusion ICU nurses demonstrates strong attitude in non-pharmacological interventions for AGI, while their knowledge and practical levels require improvement, and their KAP levels are influenced by various factors. It is recommended that nursing administrators provide systematic support, establish standardized procedures, conduct stratified training programs,and enhance ICU nurses′ satisfaction with professional growth and personal competency utilization to improve their KAP regarding non-pharmacological interventions for AGI.