MicroRNA-744 Inhibits Proliferation of Bronchial Epithelial Cells by Regulating Smad3 Pathway via Targeting Transforming Growth Factor-ß1 (TGF-ß1) in Severe Asthma.

BACKGROUND Bronchial epithelial cells proliferation plays a pivotal role in airway remodeling in children with severe asthma. MicroRNAs (miRNAs) have gained great attention for many diseases, including asthma. The purpose of this study was to explore the mechanisms that underlie miR-744 modulating bronchial epithelial cells proliferation in severe asthma in children. MATERIAL AND METHODS Bronchial epithelial cells were isolated from bronchial biopsies of normal controls and asthmatic subjects. miR-744 and transforming growth factor-ß1 (TGF-ß1) expressions were measured by quantitative reverse transcription PCR (qRT-PCR). Proliferating cell nuclear antigen (PCNA), phosphorylation or total of mothers against decapentaplegic homolog3 (Smad3) and production of Smad anchor for receptor activation (SARA) were measured via Western blot analysis. A link between miR-744 and TGF-ß1 was probed by luciferase activity and RNA immunoprecipitation. Cell proliferation was evaluated using the Cell Proliferation Assay Kit. RESULTS Severe asthma showed a significantly elevated cell proliferation rate and reduced abundance of miR-744, which in turn inhibited cell proliferation of bronchial epithelial cells. In particular, TGF-ß1 might be a candidate target of miR-744, and enrichment of miR-744 lowered the expression of TGF-ß1 at mRNA and protein levels. Indeed, overexpression of miR-744 lowered the proliferation rate of bronchial epithelial cells via driving TGF-ß1. Moreover, addition of miR-744 limited phosphorylation of Smad3 but reversed SARA protein abundance by regulating TGF-ß1. CONCLUSIONS The presence of miR-744 repressed bronchial epithelial cells proliferation through mediating the Smad3 pathway by modulating TGF-ß1, providing a promising therapeutic approach for epithelial function in severe asthma.

MFG-E8/integrin ß3 signaling contributes to airway inflammation response and airway remodeling in an ovalbumin-induced murine model of asthma.

Asthma is the most common chronic childhood disease worldwide, characterized by airway remodeling and chronic inflammation, orchestrated primarily by Th2 cytokines. The aim of the current study was to explore the influences of milk fat globule epidermal growth factor 8 (MFG-E8)/integrin ß3 signaling involved in airway inflammation and remodeling in asthma. BALB/c mice were sensitized by intraperitoneal injection of ovalbumin (OVA), followed by OVA nebulization. The levels of MFG-E8 expression were declined markedly in the OVA-induced allergy murine model. In addition, administration of MFG-E8 strongly reduced the accumulation of T-helper type 2 (Th2)-associated cytokines (such as interleukin-4, -5, and -13) as well as chemokine CCL11 (eotaxin) in bronchoalveolar lavage fluid and tissues in the OVA-sensitized mice. Moreover, MFG-E8 remarkably repressed the total immunoglobulin E and OVA-specific immunoglobulin E in serum in OVA-challenged mice. Meanwhile, treatment with recombinant murine MFG-E8 noticeably prevented inflammatory cell infiltration into the airways, as showed by a marked decrease in the numbers of total immune cells, eosinophils, neutrophils, macrophages, and lymphocytes in the bronchoalveolar lavage fluid in response to OVA challenge. Importantly, MFG-E8 apparently alleviated OVA-driven airway remodeling, which were evidenced by declined secretion of important mediators of airway remodeling, including transforming growth factor-ß1, matrix metalloproteinase 9, ADAM8, and vascular endothelial growth factor, and reduced airway collagen deposition and inhibited goblet cell hyperplasia in OVA-induced asthma in mice. Mechanistically, integrin 3 contributes to the protective effect of MFG-E8 in inhibiting airway inflammation and remodeling in OVA-driven features of allergic asthma. Overall, MFG-E8, as a candidate molecule to evaluate airway inflammation and remodeling, could be a potential target for the management and prevention of asthma exacerbations, suggesting that MFG-E8/integrin ß3 signaling may serve as a promising therapeutic agent for childhood asthma.