1,25-Dihydroxyvitamin D3 Inhibits Nuclear Factor Kappa B Activation by Stabilizing Inhibitor IκBα via mRNA Stability and Reduced Phosphorylation in Passively Sensitized Human Airway Smooth Muscle Cells

Abstract

Excessive activation of nuclear transcription factor-κB (NF-κB) is involved in human airway smooth muscle cells (HASMCs) activities in asthma. We investigated the effects of 1,25 - dihydroxyvitamin D3 [1,25 - (OH) 2D3] on the NF- κB signaling pathway in passively sensitized HASMCs and the molecular mechanisms involved. HASMCs were treated with either healthy controls' serum, asthma patients' serum or pretreated with 1,25 - (OH) 2D3 prior to treatment with asthmatics' serum. At 1 h after serum treatment: electrophoretic mobility shift assay (EMSA) was used to detect NF-κB DNA binding activity; immunocytochemical staining was used to observe the nuclear translocation of NF-κB p65; Western blots were used for NF-κB p65, IκBα, and phospho-IκBα protein levels and the nuclear translocation of NF-κB p65; real-time quantitative PCR was used for NF-κB p65 and IκBα mRNA expressions; and actinomycin D treatment was used to determine IκBα mRNA stability. Our major findings were: (1) 1,25 - (OH) 2D3 significantly reduced asthma serum passively sensitized HASMCs NF-κB DNA binding activity and inhibited the nuclear translocation of NF-κB p65; (2) 1,25 - (OH) 2D3 increased the stability of IκBα mRNA with reduced IκBα phosphorylation in asthma serum passively sensitized HASMCs and significantly increased IκBα expression in these HASMCs. Inhibiting NF-κB signalling with 1,25 - dihydroxyvitamin D3 may be a therapeutic approach for controlling HASMC-related remodelling in asthma © 2012 The Authors. Scandinavian Journal of Immunology © 2012 Blackwell Publishing Ltd.