Acute lung injury (ALI) is a severe acute inflammatory reaction of the lungs caused by a variety of factors, which can lead to a high mortality rate. MicroRNAs are a novel therapeutic molecule that play a vital role in many diseases. However, its mechanism of action in lipopolysaccharide (LPS)‐induced mouse ALI is not clear. The study aimed to investigate the mechanism of action of miR‐497 in LPS‐induced ALI. As a result, it was found that the expression of miR‐497 in the inflammatory reaction showed a decrease in time and dose trends. Importantly, miR‐497 reduced LPS‐induced expression levels of related inflammatory factors. In addition, we also demonstrated that IRAK2 is a direct target molecule of miR‐497. Interestingly, we further found that miR‐497 inhibits the expression of IRAK2 by targeting IRAK2–3′UTR. Therefore, miR‐497 can partially negatively regulate the activation of IRAK2‐NF‐κB pathway in LPS‐induced inflammatory responses.