Objective：The current study aims to investigate the relationship between hsa_circ_0005389 and neonatal acute respiratory distress syndrome（NARDS）based on high-throughput sequencing and bioinformatics analysis of human circular RNA （circRNA）to provide a new direction for the diagnosis and treatment of NARDS. Methods：The acute lung injury model was induced by lipopolysaccharide（LPS）. The expression of inflammatory markers and related pathway indexes were detected by RT-qPCR and Western blot in the negative control group and the intervention group with knockdown of hsa_circ_0005389 expression. CCK -8 and flow cytometry were used to detect the proliferation and apoptosis of blank A549 cells and A549 cells with establishment of acute lung injury model，both of which were knocked-down or over-expressed hsa_circ_0005389. Result：The mRNA relative expression and the protein expression of inflammatory markers and related pathway indexes were highly expressed and increased significantly after exposed to 10 μg/cm² LPS for 48 h（P < 0.05）. Compared with the negative control group，the relative mRNA expression level of soluble tumor necrosis factor receptor 1（sTNFR1）and the protein expression of tumor necrosis factor α（TNF-α）in A549 cells were significantly decreased（P < 0.05）after knocking down the expression of hsa_circ_0005389. Inflammatory markers and related pathway indexes decreased significantly（P < 0.001）in the acute lung injury model after knocking down the express of hsa_circ_ 0005389. The proliferation of A549 cells were accelerated and the apoptosis rate was decreased（P < 0.05）when the expression of hsa_circ_0005389 was down-regulated，while the proliferation of A549 cells was slowed down and the apoptosis rate was increased after hsa_circ_0005389 expression was over-regulated（P < 0.05）by using CCK -8 and flow cytometry as compared with the negative control group. Conclusion：In acute lung injury cell model，hsa_circ_0005389 promoted the expression of inflammatory markers of lung injury，affecting the proliferation and apoptosis of lung epithelial cells. These results suggested that hsa_circ_0005389 was involved in the inflammatory process of NARDS，which may be a potential intervention target.