Role of LncRNA H19/miR-93-5p/Orai1 axis in the regulation of human bronchial smooth muscle cell functions and airway remodeling in murine models of asthma

Abstract

\n Background\n\nType 2 cytokine Interleukin (IL)-13 regulates airway remodeling in asthma by acting on store-operated Ca2+ entry (SOCE) in airway smooth muscle cells. The underlying mechanisms of this regulating effect of IL-13 are not fully understood.\nMethods\n\nBioinformatic analysis identified interactions of microRNA 93-5p with Orai1, the pore-forming molecule of SOCE, and long non-coding RNA H19 respectively. We investigated the role of H19/miR-93-5p/Orai1 axis in the regulation of proliferation and migration of in vitro cultured human bronchial smooth muscle cells (hBSMCs) induced by IL-13. Functional relevance of H19 in airway inflammation and airway remodeling was investigated in acute and chronic murine models of asthma.\nResults\n\nIL-13 dose-dependently increased the expression of H19 and Orai1 and decreased the expression of miR-93-5p in hBSMCs; H19 siRNA reversed IL-13-induced miR-93-5p and Orai1 expression, and the proliferation and migration of hBSMCs. IL-13-promoted expression of H19 and Orai1 was reduced by miR-93-5p mimic and enhanced by miR-93-5p inhibitor. IL-13-promoted hBSMCs proliferation was enhanced by miR-93-5p inhibitor but not changed by miR-93-5p mimic; whereas IL-13-promoted hBSMCs migration was enhanced by miR-93-5p inhibitor and reduced by miR-93-5p mimic. MiR-93-5p mimic enhanced the inhibiting effect of H19 siRNA on IL-13-induced Orai1 mRNA expression, whereas miR-93-5p inhibitor reversed the inhibiting effects of H19 siRNA on IL-13-induced H19 and Orai1 mRNA and protein expression. The inhibiting effect of H19 siRNA on IL-13-induced hBSMCs proliferation and migration was reversed by miR-93-5p inhibitor but not changed by miR-935p mimic. In the lungs of both asthma mice models, the expression of H19 and Orai1 was higher than in control mice. In acute asthma mice, H19 siRNA reduced Orai1 expression, inflammatory cell infiltration and goblet cell hyperplasia in the lungs, and IL-13 levels in the bronchoalveolar lavage fluid. In chronic asthma mice, H19 siRNA reduced Orai1 expression, inflammatory cell infiltration, goblet cell hyperplasia, collagen deposition and smooth muscle mass in the lungs, as well as IL-13 levels in the BALF.\nConclusion\n\nIL-13 increases the proliferation and migration of airway smooth muscle cells by acting on H19/miR-93-5p/Orai1 axis, which also regulates airway inflammation and airway remodeling in murine models of asthma.