上海交通大学学报（医学版）

›› 2011, Vol. 31 ›› Issue (4): 397-.doi: 10.3969/j.issn.1674-8115.2011.04.003

• 论著（基础研究） • 上一篇    下一篇

机械牵张对人气道黏膜上皮细胞黏蛋白5AC表达的影响及其信号通路机制研究

钟 甜1, 尤列·皮尔曼2, 维克多·科罗索夫2, 周向东1   

  1. 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, 布拉戈维申斯克 675000
  • 出版日期:2011-04-28 发布日期:2011-04-28
  • 通讯作者: 周向东, 电子信箱: zxd999@263.net。
  • 作者简介:钟 甜（1984—）, 女, 博士生；电子信箱: crisy048@163.com。
  • 基金资助:

    国家自然科学基金（81070031）和国家自然科学基金中俄合作项目（81011120108）

Effects of mechanical stretching on expression of mucin5AC in airway epithelial cells and its mechanism of signaling pathway

ZHONG Tian1, Juliy M. Perelman2, Victor P.Kolosov2, ZHOU Xiang-dong1   

  1. 1.Department of Respiratory Medicine, The Second Affiliated Hospital, Chongqing University of Medical Science, Chongqing 400010, China；2.Far Eastern Scientific Center of Physiology and Pathology of Respiration, |Russian Academy of Medical Sciences, Blagoveschensk 675000, Russia
  • Online:2011-04-28 Published:2011-04-28
  • Supported by:

    National Natural Science Foundation of China, 81070031； Cooperation Project of China and Russia of National Natural Science Foundation of China, 81011120108

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摘要：

目的 观察机械牵张对人气道黏膜上皮细胞黏蛋白（MUC）表达的影响，并对其信号转导机制进行初步探讨。方法 采用多功能小型生物撞击机构建体外培养的人气道黏膜上皮细胞牵张模型，分别给予丝裂素活化蛋白激酶（MAPK）信号通路的三条主要通路抑制剂：JNK抑制剂（SP600125）、p38蛋白激酶抑制剂（SB203580）及ERK1/2抑制剂（U0126）（浓度均为30 μmol/L），并设未进行机械牵张的对照组和单纯机械牵张组。采用流式细胞仪检测牵张前后胞内游离Ca2+荧光强度的变化，激光共聚焦显微镜观察胞内游离Ca2+的分布情况，并分别采用RT-PCR和ELISA方法检测MUC5AC mRNA表达和蛋白分泌量。结果 牵张能显著升高人气道黏膜上皮细胞胞内Ca2+浓度、MUC5AC mRNA和蛋白表达（均P<0.01)。U0126和SB203580能抑制牵张所致MUC5AC mRNA和蛋白表达增加（均P<0.01）。结论 机械牵张能升高人气道黏膜上皮细胞黏蛋白表达，其机制可能与胞内Ca2+浓度的升高以及ERK1/2、p38 MAPK信号通路有关。

关键词: 机械牵张, 游离钙离子, 丝裂素活化蛋白激酶, 黏蛋白5AC

Abstract:

Objective To investigate the effects of mechanical stretching on the expression of mucin(MUC)5AC in airway epithelial cells, and explore its mechanism of signaling pathway. Methods Mini-type multi-fuctional bio-impact machine was used to construct the model of mechanical stretching of airway epithelial cells. Three inhibitors of mitogen-activated protein kinase (MAPK) signaling pathway (JNK inhibitor, SP600125; p38 protein kinase inhibitor, SB203580; and ERK1/2 inhibitor, U0126) were administered at the concentration of 30 μmol/L, respectively. Besides, control group without mechanical stretching and single mechanical stretching group were established. Free calcium concentrations in cells before and after mechanical stretching were determined by flow cytometry and observed by laser confocal microscopy. The expression of MUC5AC mRNA and protein was detected by RT-PCR and ELISA, respectively. Results Mechanical stretching increased the concentration of free calcium and expression of MUC5AC mRNA and protein in cells （P<0.01). U0126 and SB203580 decreased the expression of MUC5AC mRNA and protein (P<0.01). Conclusion Mechanical ventilation could increase the expression of MUC5AC in airway endothelial cells, the mechanism of which may be concerned with the increase of calcium concentration and ERK1/2 and p38 MAPK signaling pathway in cells.

Key words: mechanical stretching, free calcium ion, mitogen-activated protein kinase, mucin5AC