张力敏感性阳离子通道在机械牵张引起气道黏液高分泌中的作用  

The effects of stretch sensitive positive ion channel on the high externalization of MUCSAC in airway epithefial cells by mechanical stretching

作　　者：钟甜[1] 尤列·皮尔曼[2] 维克多·科罗索夫[2] 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,400010 [2]布拉戈维申斯克俄罗斯医学科学院远东呼吸生理与病理研究中心

出　　处：《中华医学杂志》2010年第33期2328-2333,共6页National Medical Journal of China

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）

摘　　要：目的观察张力敏感性阳离子通道、Ca^2＋内流和豆蔻酰化富丙氨酸C激酶底物（MARCKS）在机械牵张引起气道黏液高分泌中的作用。方法人气道黏膜上皮细胞（HBE16）体外培养，采用小型生物撞击机给予机械牵张刺激，各组培养细胞依施加条件不同而分为对照、牵张、牵张＋钆、牵张＋硝苯吡啶、牵张＋低分子量肝素、牵张＋MARCKS效应结构域（ED）锁核酸（LNA）以及牵张＋MARCKS的ED无关对照LNA序列共7组，分别采用逆转录聚合酶链反应（RT—Pert）和免疫荧光法观察与胞吐相关的突触相关膜蛋白SNAP23以及黏蛋白（MUC）5ACmRNA和蛋白表达，酶联免疫吸附试验（ELISA）方法检测细胞培养上清中MUC5AC分泌。结果机械牵张能显著升高人气道黏膜上皮细胞中SNAP23和MUC5AC表达，显著提升细胞培养上清中MUC5AC分泌（P〈0．05）；钆、硝苯吡啶、MARCKS的ED—LNA均能抑制机械牵张对SNAP23表达和MUC5AC表达、分泌的提升作用（均P〈0．05）；而低分子量肝素未能显著降低SNAP23表达和MUC5AC表达、分泌（P〉0．05）。结论机械牵张能升高人气道黏膜上皮细胞MUC5AC的表达，其机制可能与张力敏感性阳离子通道、Ca^2＋内流和MARCKS途径有关。Objective The effects of stretch sensitive positive ion channel, the internal flow of Ca^2＋ and myristoylated alanine-rich C kinase substrate （MARCKS） on the high externalization of MUC5AC in airway epithelial cells in mechanical stretching. Methods Mini-type Multi-fuctional Bio-impact Machin was used to construct the model of mechanical stretching. The airway epithelial cells were divide into control group, stretch, stretch and gadolinium , stretch and low molecular weight heparin, stretch and nifedipine , stretch and the locked nucleic acids of MARCKS effective domain （ ED ） , stretch and the control locked nucleic acids of MARCKS ED groups. The expression of MUCSAC and SNAP23 protein in cells were determined by immunohistochemistry method, contents of MUC5AC and SNAP23 protein were measured by Image Pro Plus 5.0. MUC5AC protein in supernatant was determined by ELISA methods. SNAP23 mRNA was determined by RT-PCR methods. Results Mechanical stretching could increase the expression of SNAP23 and MUC5AC in cells and MUCSAC in supernatant（ P 〈 0.05 ）. Gadolinium , nifedipine and LNA of MARCKS ED could reduce the increase the expression of SNAP23 and MUCSAC in cells and MUCSAC in superuatant （ all P 〈 0. 05 ）. Conclusion Mechanical stretching could increase the expression of MUC5AC in airway epithelial cells. This maybe is concerned with stretch sensitive positive ion channel, the internal flow of Ca^2＋ and MARCKS.

关 键 词：机械牵张 张力敏感性阳离子通道 胞吐 黏蛋白 

分 类 号：R363[医药卫生—病理学]