title: 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.
authors.affiliation: Department of Pulmonary and Critical Care Medicine, Fuzhou General Hospital of Nanjing Military Command, Dongfang Hospital, Xiamen University, Fuzhou, China.
authors.forename: Y
authors.initials: Y
authors.lastname: Song
authors.forename: J
authors.initials: J
authors.lastname: Hong
authors.forename: D
authors.initials: D
authors.lastname: Liu
authors.forename: Q
authors.initials: Q
authors.lastname: Lin
authors.forename: G
authors.initials: G
authors.lastname: Lai
mesh_terms: D021581:Active Transport, Cell Nucleus; D000328:Adult; D001249:Asthma / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D004247:DNA / Q000378:metabolism; D004789:Enzyme Activation / Q000187:drug effects; D005260:Female; D006801:Humans; D020672:I-kappa B Proteins / Q000235:genetics* / Q000378:metabolism*; D008297:Male; D008875:Middle Aged; D032389:Myocytes, Smooth Muscle / Q000187:drug effects* / Q000378:metabolism*; D000072000:NF-KappaB Inhibitor alpha; D016328:NF-kappa B / Q000378:metabolism*; D010766:Phosphorylation / Q000187:drug effects; D011485:Protein Binding; D020871:RNA Stability / Q000187:drug effects; D014807:Vitamin D / Q000031:analogs & derivatives* / Q000494:pharmacology; D055815:Young Adult
chemical_list: D020672:I-kappa B Proteins; D016328:NF-kappa B; C000606204:NFKBIA protein, human; C118756:dihydroxy-vitamin D3; D000072000:NF-KappaB Inhibitor alpha; D014807:Vitamin D; D004247:DNA