1,25-dihydroxyvitamin D3 inhibits nuclear factor kappa B signaling pathway in passively sensitized human airway smooth muscle cells |
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Authors: | SONG Ying-fang LAI Guo-xiang LIU De-ling LIN Qing-an LIAO Yun-hai |
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Institution: | Department of Respiratory and Critical Care Medicine, Fuzhou General Hospital of Nanjing Military Command, Fuzhou 350025, China. |
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Abstract: | AIM:To investigate the effects of 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3] on nuclear factor kappa B (NF-κB) signaling pathway in passively sensitized human airway smooth muscle cells (HASMCs). METHODS:HASMCs were passively sensitized with 10% serum from asthmatic patients. 1,25-(OH)2D3 was used as an interfering factor. Electrophoretic mobility shift assay (EMSA) was used to detect the DNA-binding activity of NF-κB. Immunocytochemical staining was used to observe the nuclear translocation of NF-κB p65. Western blotting was used for IκBα and phosphorylated IκBα protein detection. Real-time fluorescence quantitative PCR was used to determine vitamin D receptor (VDR), vitamin D 24-hydroxylase (CYP24) and IκBα mRNA expression. The mRNA expression of IκBα in HASMCs after actinomycin D treatment was also determined. RESULTS:(1) 1,25-(OH)2D3 significantly attenuated the DNA-binding activity of NF-κB and the nuclear translocation of NF-κB p65 in HASMCs passively sensitized by asthmatic serum. (2) 1,25-(OH)2D3 enhanced IκBα mRNA stability and inhibited IκBα protein phosphorylation in passively sensitized HASMCs, thus increasing IκBα expression in these HASMCs. (3) 1,25-(OH)2D3 up-regulated VDR mRNA level and evoked its functional response in passively sensitized HASMCs. CONCLUSION: 1,25-(OH)2D3 enhanced the expression of IκBα and therefore inhibited NF-κB signaling passway in HASMCs. This effect may be dependent on VDR, and responsible for the inhibitory effect of 1,25-(OH)2D3 on passively sensitized HASMCs. |
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Keywords: | Asthma Airway smooth muscle cells 1 25-Dihydroxyvitamin D3 Nuclear factor kappa B Receptors vitamin D |
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