Objective To observe the effect of audio-visual-tactile-olfactory interventions on neonatal venipuncture pain, and explore effective methods to reduce neonatal venipuncture pain. Methods A total of 90 cases with neonatal pneumonia were randomly divided into the control group and the observation group, with 45 cases in each group. The control group received routine nursing care for venipuncture, and the observation group received audio-visual-tactile-olfactory interventions of wearing an eye mask, listening to their mother′s voice and heartbeat, touch, and dripping 24% sucrose water on their anterior tongue on the basis of routine nursing care. The neonatal pain score, physiological parameters, and the success rate of one-time venipuncture were compared between the two groups. Results The neonatal pain score during puncture in the observation group was significantly lower than that in the control group, the heart rate, respiration, and blood oxygen saturation were significantly better than those in the control group (all P<0.05). The success rate of one-time venipuncture in the control group was higher than that in the control group, but the difference between the two groups was not statistically significant (P>0.05). Conclusion Audio-visual-tactile-olfactory interventions can effectively reduce neonatal puncture pain and maintain their physiological parameters stability.