A dynamic real-time method for monitoring epithelial barrier function in vitro
A dynamic real-time method for monitoring epithelial barrier function in vitro
复制标题
体外动态实时监测上皮屏障功能的方法
DOI：
10.1016/j.ab.2012.03.010
复制
发表时间：
2012-06-15
影响因子：
2.9
通讯作者：
Ke, Yuehai
中科院分区：
文献类型：
--
作者：
Sun, Mingjiao;Fu, Huiying;Ke, Yuehai
肠上皮作为抵抗有害的内腔环境的物理屏障发挥作用，其通常在存在肠道疾病时受损。在这项工作中，我们引入了一个电子阻抗为基础的分析，使用实时xCELLigence系统记录乙醇诱导的肠道屏障功能障碍的动态过程。在分析形态学改变的细胞旁连接复合体和组织的细胞周围的F-肌动蛋白，这种新的，实时的，基于细胞的技术显示出相当大的相关性与标准的跨上皮电阻终点测定。此外，在真实的时间内监测屏障功能允许无偏地筛选和表征管腔中影响上皮完整性的生化剂。该功能测定进一步鉴定了诱导型一氧化氮合酶抑制剂、上皮生长因子、酪氨酸激酶和磷酸酶在响应乙醇给药调节上皮屏障功能中的体外作用。总之，我们的研究结果表明，这种新的，实时的，高通量的方法提供了一个有前途的工具，监测上皮屏障功能的情况下，更多的生理相关性。(C)2012 Elsevier Inc. All rights reserved.
The intestinal epithelium functions as a physical barrier against the harmful environment of the lumen, which usually becomes impaired in the presence of intestinal diseases. In this work, we introduce an electronic impedance-based analysis using a real-time xCELLigence system to record the dynamic processes of ethanol-induced intestinal barrier dysfunction. In terms of analyzing morphological alterations in the paracellular junction complex and the organization of pericellular F-actin, this novel, real-time, cell-based technology shows considerable correlations with the standard transepithelial electrical resistance endpoint assay. In addition, monitoring barrier functions in real time allows unbiased screening and characterization of biochemical agents in the lumen that affect epithelial integrity. This functional assay further identifies the in vitro roles of the inducible nitric oxide synthase inhibitor, epithelial growth factor, tyrosine kinases, and phosphatases in regulating epithelial barrier function in response to ethanol administration. Taken together, our findings suggest that this novel, real-time, high-throughput method offers a promising tool for monitoring epithelial barrier functions in situations with more physiological relevance. (C) 2012 Elsevier Inc. All rights reserved.