Acute respiratory distress syndrome (ARDS) is a critical condition with high mortality. High mobility group protein B1 (HMGB1) is one of the key proinflammatory factors in the ARDS "inflammatory storm". According to previous studies, some miRNAs play important roles in this process. We aimed to determine the contributing miRNAs targeting the expression and release of HMGB1. MiRNA expression in the peripheral blood of patients with ARDS was measured by microRNA microarray. MiRNAs targeting HMGB1 was screened and explored for further study. In LPS-induced cell and mouse ARDS models, we explored the effect of this miRNA on the expression and secretion of HMGB1 by western blot (WB), real-time quantitative polymerase chain reaction (RT-qPCR) and enzyme-linked immunosorbent assay (ELISA). The effects of this miRNA on the nuclear factor κB (NF-κB) signaling pathway, proinflammatory cytokines and nod-like receptor protein 3 (NLRP3) inflammasome were detected by WB and RT-qPCR. In ARDS models, MiR-574-5p expression could be induced by the TLR4/NF-κB pathway upon LPS stimulation. It could suppress the inflammatory response by targeting HMGB1. Enforced the expression of miR-574-5p or HMGB1 siRNA silencing inhibits the activation of NF-κB signaling pathway and the NLRP3 inflammasome. Moreover, overexpression of HMGB1 reversed the anti-inflammatory effect of miR-574-5p. In ARDS mice, overexpression of miR-574-5p suppresses the alveolar leukocytes infiltration, interstitial edema, protein effusion and inflammation. This study demonstrated that miR-574-5p provided negative feedback to LPS-induced inflammation and relieved ARDS. It may provide new therapeutic strategies for ARDS.