高渗诱导人气道上皮细胞系黏液高分泌  

Hypertonicity induces mucus hypersecretion by human airway epithelial cells

作　　者：涂静[1] 尤列.皮尔曼 维克多.科罗索夫 周向东[1] 

机构地区：[1]重庆医科大学附属第二医院呼吸内科,重庆400010 [2]俄罗斯医学科学院远东呼吸生理与病理研究中心,俄罗斯布拉戈维申斯克675000

出　　处：《基础医学与临床》2011年第7期723-727,共5页Basic and Clinical Medicine

基　　金：国家自然科学基金(30770951);国家自然科学基金中俄合作项目(81011120108);中俄政府间合作项目(20091301)

摘　　要：目的研究高渗条件对正常人气道上皮细胞(HBE)黏蛋白(MUC)5AC分泌的影响,以及蛋白激酶C(PKC)-热休克蛋白(HSP)70信号途径在其中的可能作用。方法 采用高渗盐水诱导培养HBE16细胞的方法复制黏液高分泌体外模型,分别用PKCμ抑制剂G 6976、PKCα抑制剂Safingol、PKCβ抑制剂LY333531和PKCδ抑制剂Rot-tlerin干预HBE16细胞。Western blot检测HSP70-2的蛋白含量;RT-PCR检测人HSP70-2转录水平;ELISA检测培养上清MUC5AC蛋白含量c各高渗组的培养上清MUC5AC蛋白含量、HSP70-2蛋白和转录水平较对照组显著升高,并随着培养时间的延长而逐渐增加(P<0.05)。G 6976处理组的上述指标显著降低(P<0.01),但仍高于对照组(P<0.05);而Safingol、LY333531和Rottlerin处理组均无改变。结论 高渗盐水可诱导人气道上皮细胞MUC5AC的高分泌,HSP70-2系通过PKC中的μ亚型在该过程中起重要作用的。Objective To investigate the effect of hypertonic conditions induces mucin（MUC）5AC secretion by HBE16 cell and the potential mechanism of PKC-HSP70 pathways.Methods HBE16 cells were cultured in hypertonic medium to induce mucus hypersecretion in vitro.Inhibitors of PKC isoforms were used to interference HBE16 cells.HSP70-2 cells were determined by Western blot.RT-PCR was used to detecte the transcriptional level of HSP70-2.Mucin（MUC）5AC protein content in supernatant was analysed by ELISA.Results The levels of HSP70-2,HSP70-2 mRNA and MUC5AC proteins werer strongly increased after cells were exposed to hypertonic conditions,and the expression content was increased in a time-dependent manner（all P0.05）.The expression in G6976 group was significantly lower than that in hypertonic group（P0.01） and was still higher than that in control（P0.05）.Treatment with Safingol,LY333531 and Rottlerin exerted no effects on hypertonicity-induced content of HSP70-2,HSP70-2 mRNA and MUC5AC proteins.Conclusion Hypertonic medium induces the MUC5ACmucin hypersecretion by human bronchial epithelial cells.HSP70-2 mediated hypertonic stress induces mucin hypersecretion through a PKCμ dependent signaling pathways.

关 键 词：热休克蛋白70 黏蛋白 渗透压 蛋白激酶C 

分 类 号：R562[医药卫生—呼吸系统] R363.1[医药卫生—内科学]