MicroRNA-21 released from mast cells-derived extracellular vesicles drives asthma in mice by potentiating airway inflammation and oxidative stress

Objective Mast cells-derived extracellular vesicles (EVs) play vital roles in various physiological and pathophysiological conditions. However, the cargoes of mast cells-derived EVs in asthma have not been established. Here, we set to identify the role of microRNA-21 (miRNA-21) from mast cells-derived EVs in ozone- and lipopolysaccharide (LPS)-induced mouse airway epithelial cells (MIC-iCell-a006 cells) and asthmatic mice. Methods After ozone or LPS treatment, MIC-iCell-a006 cells were subjected to a microarray analysis to screen differentially expressed miRNAs, and then co-cultured with EVs. miR-21 was silenced in cells, followed by CCK-8, scratch, and Transwell assays. Mice were challenged with ovalbumin, and antioxidant enzymes and inflammatory cell infiltration were assessed after EVs and miR-21 inhibitor treatments. The relation between miR-21 and DDAH1 was evaluated by Dual-luciferase assay, and changes in Wnt/β-catenin pathway related proteins were examined by western blot. Finally, the involvement of the DDAH1/Wnt/β-catenin axis in miR-21-mediated oxidative stress and inflammation was verified by rescue experiments. Results miR-21 expression was upregulated in MIC-iCell-a006 cells induced by ozone or LPS. miR-21 was enriched in mast cells-derived EVs, and EVs increased miR-21 expression in MIC-iCell-a006 cells. miR-21 inhibitor increased cell activity and alleviated oxidative stress and inflammation. In asthmatic mice, miR-21 expression was increased, and EVs decreased antioxidant enzymes and increased inflammatory cells, whose effects were reversed by miR-21 knockdown. miR-21 targeted DDAH1 to mediate the Wnt/β-catenin signaling, and down-regulation of DDAH1 inhibited the action of miR-21 inhibitor. Conclusion The miR-21 secreted from mast cells-derived EVs promotes oxidative stress and inflammatory responses in asthmatic mice via the DDAH1/Wnt/β-catenin signaling axis.