Objective: To gain a new direction about distinguishing and treating neonatal ARDS, the relationship between hsa_circ_0005389 and neonatal ARDS, based on human circRNA high-throughput sequencing and the results of bioinformatics analysis, was studied. Methods: The acute lung injury model was induced by lipopolysaccharide. The expression of inflammatory markers were detected by qRT-PCR and Western blot in the A549 cell knocked down the expression of hsa_circ_0005389 and (or no) exposure LPS. And then the proliferation and apoptosis of A549 cells knocked down the expression of hsa_circ_0005389 were detected by CCK8 and flow cytometer. Result: The mRNA relative expression and the protein expression of all the research indexes increased significantly in the group of 10 μg/cm2 LPS(P<0.0001). Compare with the black and negative control (NC) group, it is decreased significantly that the mRNA relative expression of sTNFR1 and the protein expression of TNFα in A549 cells knocked down the expression of hsa_circ_0005389 (P<0.05). But all the research indexes decreased significantly (P<0.0001) in the group which knocked down the expression of hsa_circ_0005389 and then exposed LPS. In addition, the results of CCK8 showed that the proliferation of si-hsa_circ_0005389 group, compared with other groups, was different after 48 hours and 72 hours of culture (P<0.05). Compared with the apoptosis rate of the NC group, the apoptosis rate of the si-hsa_circ_0005389 group decreased significantly. Conclusion: In the acute lung injury model, hsa_circ_0005389 promotes the expression of inflammatory factors, such as TNFR1,TNFα and so on. At same time, it inhibits the proliferation of A549 and boosts the apoptosis. So hsa_circ_0005389 accelerates the inflammatory process of neonatal ARDS.