To analyze the application effect of caffeine citrate combined with pulmonary surfactant (PS) and non-invasive ventilator for treating premature infants with respiratory distress syndrome (RDS). Methods: The clinical data of the premature infants with RDS admitted to the hospital between July 2021 and July 2023 were analyzed retrospectively. 45 infants who had received the treatment of PS combined with nasal continuous positive airway pressure (nCPAP) were included in the study group, and 40 infants who had received the treatment of the caffeine citrate combined with PS and nCPAP were selected in the study group. The duration of the hospital stay, the oxygen therapy time, the apnea frequency and the complications occurrence of the infants were compared between the two groups. The changes of the blood gas indicators and the serum inflammatory factors of the infants in the two groups were analyzed. The clinical efficacy of the infants after 3 days of treatment was compared between the two groups. Results: After treatment, the hospitalization time (23.12±2.03d), the total oxygen used time (80.14±20.58h) and the apnea number (4.02±0.58 times) of the infants in the study group were significantly lower than those (25.07±2.69d, 106.43±32.61h and 7.14±1.03 times) of the infants in the control group. The pH value and PaO2 level of the infants in the two groups after treatment had increased significantly, and which (7.35±0.10 and 69.54±9.02 mmHg) of the infants in the study group were significantly higher than those (7.20±0.05 and 60.37±8.14 mmHg) of the infants in the control group. The PaCO2 value and the serum inflammatory factors levels of the infants in the two groups after treatment had decreased significantly, and the PaCO2 value (240.16±5.63 mmHg) of the infants in the study group was significantly lower than that (43.08±6.31 mmHg) of the infants in the control group (all P<0.05). After 3 days of treatment, the total effective rate (87.5%) of the infants in the study group was significantly higher than that (68.9%) of the infants in the control group, and the incidence (5.0%) of the bronchopulmonary dysplasia (BPD) of the infants during the treatment was significantly lower than that (22.2%) of the infants in the control group (all P<0.05). Conclusion: The caffeine citrate combined with PS and nCPAP therapy for the premature infants with RDS can effectively shorten their number of apnea and oxygen used time within 24h after born, improve their blood gas indicators and body’ s inflammatory response, reduce the incidence of their BPD and promote their rehabilitation.