畜牧兽医学报

• 基础兽医 • 上一篇    下一篇

细胞信号转导和骨架在人源福氏志贺菌侵袭鸡肠上皮原代细胞中的作用

师润,杨霞,陈陆*,刘红英,常洪涛,余秋颖,明星,郑关民,王川庆   

  1. (河南农业大学牧医工程学院,郑州 450002)
  • 收稿日期:2012-07-25 出版日期:2013-02-23 发布日期:2013-02-23
  • 通讯作者: 陈陆,男,河南信阳人,副教授,博士,主要从事动物传染病发病机制及免疫防治研究,E-mail:chenluhau@126.com
  • 作者简介:师润(1986-),男,河南平顶山人,硕士生,主要从事动物疫病发病机制及免疫防治研究,E-mail:shirunok.2006@163.com
  • 基金资助:

    国家自然科学基金(30972187);教育部高等学校博士学科点专项科研基金(2011410511003)

The Role of Cell Signal Transduction and Cytoskeleton in Invasion of Primary Chicken Intestinal Epithelial Cell by Human Source S. flexneri

SHI Run, YANG Xia, CHEN Lu*, LIU Hong-ying, CHANG Hong-tao, YU Qiu-ying, MING Xing, ZHENG Guan-min, WANG Chuan-qing   

  1. (College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450002, China)
  • Received:2012-07-25 Online:2013-02-23 Published:2013-02-23

摘要:

为研究细胞信号转导和骨架在人源福氏志贺菌侵袭鸡肠上皮原代细胞中的作用,分别用酪氨酸蛋白激酶、酪氨酸蛋白磷酸酶、蛋白激酶C、Ca2+通道等信号转导抑制剂染料木黄酮、原钒酸钠、星状孢子素、硝苯地平和微丝蛋白聚合抑制剂细胞松弛素B处理鸡肠上皮原代细胞后,采用细胞免疫组织化学染色检测人源福氏志贺菌ZD02株对鸡肠上皮原代细胞侵袭率,用间接免疫荧光双重标记检测ZD02株侵袭鸡肠上皮原代细胞后F-肌动蛋白分布的变化。结果显示,染料木黄酮、硝苯地平和细胞松弛素B能显著抑制人源福氏志贺菌ZD02株内化,星状孢子素和原钒酸钠对ZD02株的内化无影响,人源福氏志贺菌侵袭鸡肠上皮原代细胞时F-肌动蛋白发生聚集。本研究表明人源福氏志贺菌ZD02株侵袭鸡肠上皮原代细胞过程中需要酪氨酸蛋白激酶、Ca2+通道活化以及细胞微丝重排。

Abstract:

This experiment was conducted to study the role of cell signal transduction and cytoskeleton in invasion of primary chicken intestinal epithelial cell by human source S. flexneri. Primary chicken intestinal epithelial cell was treated with Genistein, sodium orthovanadate, staurosporine, Nifedipine and Cytochalasin B respectively, which are the corresponding inhibitor of protein tyrosine kinase, protein tyrosine phosphorylase, protein kinase C, Ca2+ channel signal transduction and microfilament polymerization. The influences of above inhibitors on invasion of primary chicken intestinal epithelial cells by ZD02 strain were detected by immunohistochemistry. F-actin distribution of primary chicken intestinal epithelial cells infected with Human S. flexneri ZD02 strain was detected by indirect immunofluorescence double labeling. Results were as follows: Genistein and Nifedipine significantly reduced internalization of ZD02 strain into primary chicken intestinal epithelial cells. In contrast, staurosporine and sodium orthovanadate have no effect on internalization of ZD02 strain. Cytochalasin B could prevent internalization of ZD02 strain. F-actin aggregated when ZD02 strain invaded into primary chicken intestinal epithelial cells. The process of human source S. flexneri invading into primary chicken intestinal epithelial cell depends on protein tyrosine kinase, Ca2+ channel activation and actin rearrangement.

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