Abstract

 
 
 
 Purpose: To investigate the action of miR-483-3p in pediatric pneumonia and identify potential biomarkers.
 Methods: Bronchoalveolar lavage was collected from 38 pneumonia patients and 25 healthy children. The expression of miR-483-3p in the lavage was determined using qRT-PCR. Alveolar macrophages collected from lavage of healthy children were cultured and used for functional assays. Transwell assay was conducted to evaluate macrophage cell migration. Cell viability and apoptosis were evaluated in lipopolysaccharide (LPS)-induced human pulmonary alveolar epithelial cells (HPAEpiCs) by CCK8 (cell counting kit - 8) or flow cytometry, respectively.
 Results: MiR-483-3p was significantly elevated in bronchoalveolar lavage of pneumonia patients, when compared to healthy children (p < 0.001). MiR-483-3p, which targets insulin-like growth factor 1 (IGF1), decreased the mRNA and protein expression of IGF1 in alveolar macrophages collected from the lavage of healthy children. MiR-483-3p reduced motility of macrophages. IGF1 counteracted the LPS- induced decrease in cell viability and the increase in apoptosis of HPAEpiCs. Conditioned medium from macrophages transfected with miR-483-3p inhibitor increased cell viability and reduced cellular apoptosis of LPS-induced HPAEpiCs. However, conditioned medium from macrophages transfected with miR-483-3p mimics decreased cell viability and increased apoptosis.
 Conclusion: MiR-483-3p negatively regulates IGF1 to promote progression of pediatric pneumonia, providing a potential therapeutic target in pediatric pneumonia.
 
 
 
Highlights
Pneumonia, a common respiratory disease caused by pathogens or viral infection, is the leading cause of mortality and morbidity in children [1]
Insulin-like growth factor 1 levels in alveolar macrophage culture medium were determined via insulin-like growth factor 1 (IGF1) antibody DuoSet (R&D Systems, Minneapolis, MN, USA), following the manufacturer’ s instructions
Conditioned medium from macrophages transfected with miR483-3p mimics aggravated LPS-induced cell injury of HPAEpiC, confirming that miR-483-3p might contribute to pediatric pneumonia
Summary
A common respiratory disease caused by pathogens or viral infection, is the leading cause of mortality and morbidity in children [1]. Research has shown that macrophages secrete cytokines to participate in alveolar epithelial cell injury during pneumonia [4]. Alveolar macrophage-derived IGF1 may be an effective target for the treatment of pediatric pneumonia. Bronchoalveolar lavage cells of healthy children were suspended in RPMI-1640 medium (Lonza, Basel, Switzerland) containing 10 % fetal bovine serum (Gibco, Waltham, MA, USA). Insulin-like growth factor 1 levels in alveolar macrophage culture medium were determined via IGF1 antibody DuoSet (R&D Systems, Minneapolis, MN, USA), following the manufacturer’ s instructions. Alveolar macrophages suspended in RPMI 1640 medium were placed in the upper compartment of vitronectin-coated chambers (BD Biosciences, Bedford, MA, USA). Samples of conditioned medium from macrophages transfected with miR-483-3p mimics or inhibitor were collected via centrifugation at 2500 rpm for 30 min. Values of p < 0.05 were considered statistically significant
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