Objective To explore the effect of non nutritive sucking combined with sucrose solution on alleviating pain response during venous puncture in premature infants,providing a convenient and effective analgesic strategy for impro-ving clinical care of premature infants.Methods A total of 114 premature infants admitted to the neonatal department of Children's Hospital of Nanjing Medical University from October 2021 to October 2022 were selected and randomly divided into two groups using a random number table method,with 57 cases in each group.Both groups were subjected to venous puncture for blood collection,while the control group was subjected to non nutritive sucking intervention before puncture,and the study group was combined with sucrose solution intervention on this basis.The changes in pain degree,heart rate,blood oxygen saturation,salivary catecholamine concentration and salivary cortisol concentration before and after puncture were compared between the two groups.Results The pain score during puncture in the study group was(5.67±1.02)points,which was lower than that of the control group[(9.73±1.28)points,P<0.05].The heart rates of the study group during puncture and 5 minutes after puncture were(150.74±5.16)beats/min and(145.17±7.29)beats/min,respectively,which were lower than those of the control group[(165.09±5.33)beats/min and(160.75±9.36)beats/min,P<0.05].The blood oxygen saturation during puncture and 5 minutes after puncture in the study group were(94.40±0.23)%and(94.48±0.37)%,respectively,which were higher than those in the control group[(94.02±0.44)%and(94.16±0.25)%,P<0.05].There was no statistically significant difference in the concentra-tion of salivary catecholamines or salivary cortisol between the study group and the control group at the time of puncture,5 minutes after puncture,and 20 minutes after puncture(P>0.05).Conclusion Sucrose solution combined with non nutritive sucking effectively alleviate the pain response to venous puncture in premature infants and stabilize the vital signs.