基础医学与临床 ›› 2016, Vol. 36 ›› Issue (11): 1483-1487.

• 研究论文 • 上一篇    下一篇

BRAP对LPS诱导的人气道上皮细胞株黏液高分泌的影响

胥青,周向东   

  1. 重庆医科大学附属第二医院 呼吸内科
  • 收稿日期:2015-12-22 修回日期:2016-03-31 出版日期:2016-11-05 发布日期:2016-10-24
  • 通讯作者: 周向东 E-mail:zxd999@263.net
  • 基金资助:
    拴链因子Exocyst复合物在慢性气道炎症中的促黏液分泌效应

Effect of BRAP on mucous hypersecretion induced by LPS in human airway epithelial cells

  • Received:2015-12-22 Revised:2016-03-31 Online:2016-11-05 Published:2016-10-24
  • Contact: Xiang-dong Zhou E-mail:zxd999@263.net

摘要: 目的 探讨蛙皮素受体激活蛋白(BRAP)对脂多糖(LPS)诱导的气道黏液高分泌的影响及相关作用机制。方法 体外培养人气道上皮HBE16细胞,转染已构建好的pEGFP-N1-BRAP,以空质粒载体作为对照,给予LPS刺激,同时分别以ERK抑制剂U0126、JNK抑制剂SP600125、p38抑制剂SB203580干预。观察细胞内活性氧(ROS)含量、黏蛋白(MUC)5AC表达和核因子-ΚB(NF-ΚB)活性以及细胞活力的变化。结果 转染pEGFP-N1-BRAP后ROS明显减少(P < 0.01);同单纯LPS刺激相比,MUC5AC蛋白含量和mRNA水平在转染pEGFP-N1-BRAP后明显降低(P < 0.01),NF-ΚB活性亦呈相同趋势(P < 0.05);在U0126组,MUC5AC的表达较转染重组质粒组显著降低(P < 0.01),NF-ΚB活性也显著下调(P < 0.05)。结论 BRAP可以通过ROS/ERK/MAPK信号通路阻止胞质内NF-ΚB的活化,从而下调MUC5AC的生成。

关键词: 关键词 蛙皮素受体激活蛋白, 黏蛋白5AC, 细胞外信号调节蛋白激酶, 核转录因子-ΚB

Abstract: Objective To investigate the effect of bombesin receptor activated protein(BRAP) on lipopolysaccharide(LPS)-induced mucous hypersecretion in airway and mechamism. Methods After cultivating HBE16 cells in vitro , the constructed p-EGFP-N1-BRAP eukaryotic vector was transfected into the cells,with empty plasmid as a vector control. At the same time in order to LPS as a stimulus to extracellular signal-regulated protein kinase(ERK) specific inhibitor U0126,P38 inhibitor SB203580 and c-jun N-terminal kinase (JNK) inhibitor SP600125 for the intervention factors. The content of ROS, the expression of MUC5AC mRNA and protein, the NF-ΚB activity and cell vitality were detected. Results After transfected pEGFP-N1-BRAP, ROS production was decreased (P < 0.01). The MUC5AC protein production and mRNA level of the group transfected pEGFP-N1-BRAP were significantly decreased as compared with LPS group(P < 0.01), the NF-ΚB activity showed the same trend(P < 0.05). In U0126 group, the expression of MUC5AC was significantly decreased as compared with transfected recombination plasmid group(P < 0.01), the NF-ΚB activity was also significantly decreased(P < 0.05). Conclusions BRAP prevents NF-ΚB activation via ROS/ERK/MAPK signaling pathway,thus reducing LPS-induced mucin5AC production.

Key words: Key words: bombesin receptor activated protein, mucin5AC, extracellular signal-regulated protein kinase, nuclear factor-ΚB

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