1,25-二羟维生素D_3负性调控被动致敏人气道平滑肌细胞中的NF-κB信号通路  

1,25-dihydroxyvitamin D_3 inhibits nuclear factor kappa B signaling pathway in passively sensitized human airway smooth muscle cells

作　　者：宋颖芳[1] 赖国祥[1] 柳德灵[1] 林庆安[1] 廖云海[1] 

机构地区：[1]南京军区福州总医院呼吸与危重症医学科,福建福州350025

出　　处：《中国病理生理杂志》2013年第11期2044-2048,共5页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81100011);福建省自然科学基金资助项目(No.2011J05087)

摘　　要：目的:探讨1,25-二羟维生素D3[1,25-(OH)2D3]对被动致敏人气道平滑肌细胞(HASMCs)中核因子κB(NF-κB)信号通路的影响。方法:原代培养HASMCs并使之被动致敏,以1,25-(OH)2D3作为干预因素。EMSA法检测NF-κB的DNA结合活性;免疫细胞化学染色技术观察NF-κB p65的核易位情况;Western blotting法检测核因子κB抑制蛋白α(IκBα)及p-IκBα蛋白的表达水平;实时荧光定量PCR检测维生素D受体(VDR)、维生素D 24-羟化酶(CYP24)和IκBαmRNA的表达水平;放线菌素D处理实验检测IκBαmRNA的表达。结果:(1)1,25-(OH)2D3显著削弱被动致敏HASMCs中NF-κB的DNA结合活性及其亚单位p65的核易位;(2)1,25-(OH)2D3能通过增加被动致敏HASMCs中IκBα的mRNA稳定性及减少其蛋白磷酸化水平2个途径显著上调细胞中IκBα的表达;(3)1,25-(OH)2D3显著上调被动致敏HASMCs中VDR的mRNA表达并诱发其功能性反应。结论:1,25-(OH)2D3能通过上调被动致敏HASMCs中IκBα的表达抑制细胞NF-κB信号通路,且这一作用与VDR有关,这可能是其调控被动致敏HASMCs的重要作用机制。AIM： To investigate the effects of 1,25-dihydroxyvitamin D3 [ 1,25-（ OH ） 2 D3 ] 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）2 D3 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 IκBa 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） 2 D3 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）：D3 enhanced IκBα mRNA stability and inhibited IκBoL protein phosphorylation in passively sensitized HASMCs, thus increasing IκBa 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） 2 D3 on passively sensitized HASMCs.

关 键 词：哮喘 气道平滑肌细胞 1 25-二羟维生素D3 核因子κβ 受体 维生素D 

分 类 号：R363[医药卫生—病理学]