基础医学与临床 ›› 2009, Vol. 29 ›› Issue (11): 1139-1143.

• 研究论文 • 上一篇    下一篇

TNF-α与香烟烟雾在诱导气道上皮细胞株黏蛋白表达的相互作用

李琪 周向东   

  1. 重庆医科大学 第二附属医院 呼吸内科 重庆医科大学 第二附属医院 呼吸内科
  • 收稿日期:2009-03-02 修回日期:2009-03-30 出版日期:2009-11-20 发布日期:2009-11-20
  • 通讯作者: 周向东

Interaction of TNF-α and cigarette smoke extract in respiratory mucus expression

Qi LI, Xiang-dong ZHOU   

  1. Department of Respiratory Medicine,the Second Affiliated Hospital,Chongqing Medical University Department of Respiratory Medicine,the Second Affiliated Hospital,Chongqing Medical University
  • Received:2009-03-02 Revised:2009-03-30 Online:2009-11-20 Published:2009-11-20
  • Contact: Xiang-dong ZHOU

摘要: 目的 探讨肿瘤坏死因子TNF-α与香烟提取物共同诱导气道黏液高分泌的相互关系及作用特点。方法 培养的人气道上皮细胞BEAS-2B,转染核转录因子(NF)-κB"decoy"寡核苷酸(ODNs),并以乱序NF-κB ODNs为对照转染组,各组均分别予以TNF-α、10%香烟提取物(CSE)单独刺激及共同刺激,以Western blot、ELISA及RT-PCR法检测各组刺激前后磷酸化表皮生长因子受体(p-EGFR)、黏蛋白(MUC5AC)蛋白及mRNA水平。结果 转染乱序NF-κB ODNs的细胞予以TNF-α、10% CSE刺激后,各组细胞中p-EGFR蛋白水平较对照组升高,伴随MUC5AC蛋白含量及基因转录水平的提高(P<0.05);共孵育组中较单独刺激组升高更为显著(P<0.05)。转染NF-κB"decoy"ODNs组再予以TNF-α刺激,细胞中MUC5AC蛋白含量及mRNA水平与未转染组相比升高不明显,而单用CSE刺激组中的MUC5AC、p-EGFR水平仍有明显升高(P<0.05);共孵育组显示出与CSE单独刺激组相似的结果。结论 TNF-α、CSE能协同促进气道上皮细胞中黏蛋白合成,转录因子NF-κB主要参与TNF-α所致的MUC5AC表达,而在CSE诱导的效应过程中作用不明显。

关键词: 黏蛋白类, 肿瘤坏死因子α, 吸烟, 核转录因子-κB

Abstract: Objective To explore the mutual effect of proinflammatory factor tumor necrosis factor(TNF)-α and cigarette smoke extract(CSE) in the induction of mucin 5AC in BEAS-2B human airway epithelial cells.Methods The BEAS-2B cells were transfected with nuclear factor(NF)-κB decoy oligonucleotides(ODNs) or transfected with NF-κB scrambled ODNs as negative control,then treated with TNF-α,CSE,and TNF-α plus CSE,respectively. The expression of phosphorylated epidermal growth factor receptor(p-EGFR),mucin 5AC peotein production and gene expression were detected by Western blot,ELISA and RT-PCR,respectively.Results It was showed an obvious increasing of p-EGFR protein production in scrambled ODNs transfected cells exposed to stimuli,with elevation of MUC5AC protein production and mRNA expression,and the TNF-α and CSE co-incubated groups were much higher than single TNF-α or CSE stimulated group(P<0.05).The relative content of p-EGFR protein,MUC5AC mRNA,and MUC5AC protein in the TNF-α and CSE coincubated groups were 0.74±0.06,0.77±0.10,and 0.72±0.03,respectively,compared with 0.58±0.08,0.52±0.07,and 0.53±0.11 in CSE-stimulated group and 0.53±0.04,0.58±0.11,0.56±0.05 in TNF-α-induced group,all P<0.05. In NF-κB decoy ODNs transfected cells,TNF-α stimuli revealed an obvious reduction of MUC5AC protein production and gene expression,were 0.35±0.12,and 0.41±0.12,respectively,compared with scrambled ODNs transfected group,P<0.05,but had little effect in CSE group and co-incubated group,which still at a relative high level, were 0.48±0.04,0.49±0.11 and 0.51±0.08,0.48±0.07,respectively,compared with negative control group,P<0.05. Conclusion TNF-α and CSE can synergistically induce mucin 5AC expression in BEAS-2B cells, transcription factor NF-κB plays a critical role in TNF-α induced mucin production but not as so important in CSE stimulated cells.

Key words: Mucins, tumor necrosis factor-alpha, cigarette smoking, NF-kappa B

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