Objective：To investigate the role of NANCI-NKX2.1 signaling pathway in the process of bronchopulmonary dysplasia（BPD） formation in neonatal mice. Methods：A total of 32 neonatal mice（C57BL/6J） were randomly divided into 4 groups：interference group，negative control group，normal saline control group and blank control group，with 8 pups in each group. One microlitre of normal saline with lncRNA NANCI adenovirus and control adenovirus were administered intranasally to lungs of newborn mice in interference group and negative control group at postnatal days 2 respectively. One microlitre of normal saline was administered intranasally to lungs of newborn mice in normal saline control group .No interference treatment was processed in blank control group．The mice were sacrificed 7 days after birth and lung tissue samples were collected. Hematoxylin-eosin（HE）staining was used to observe pathological changes in lung tissues. RT-qPCR was used to measure the expression of NANCI. RT-qPCR and Western blot were used to measure the mRNA and protein expression of NKX2.1，SPC and AQP5. Results：HE staining in interference group showed thinner alveolar septum，increased alveolar cavity and decreased number of alveoli and septum. Lung tissue biopsy also showed decreased RAC values（P < 0.05）.The relative expression of NANCI decreased（P < 0.05）；the expression of NKX2.1，SPC and AQP5 mRNA and protein were decreased（P < 0.05）. Conclusion：NANCI-NKX2.1 signaling pathway may play an important role in BPD neonatal mice.