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-κBODNs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-κBODNs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"de-coy"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。
Objective To explore the mutual effect of proinflammatory factor tumor necrosis factor (TNF)-α and cigarette smoke extract(CSE) in the induction of mucin SAC 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 5 AC ( MUC5AC ) protein production and gene expression were detected by Western blot, ELISA and RT-PCR respectively. Results Significant increasing of p-EGFR protein production in scrambled ODNs transfected cells with elevation of MUCSAC protein production and mRNA expression was recorded and TNF-α and CSE co-incubated groups were much higher than single TNF-α or CSE stimulated group( P 〈 0. 05 ). In NF-κB decoy ODNs transfected cells ,TNF-α stimuli revealed an obvious reduction of MUCSAC protein production and gene expression. Conclusion TNF-α and CSE can syner- gistically 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 mucin production by CSE stimulated cells.
出处
《基础医学与临床》
CSCD
北大核心
2009年第11期1139-1143,共5页
Basic and Clinical Medicine
基金
国家自然科学基金(30770951)