title: A dynamic real-time method for monitoring epithelial barrier function in vitro.
abstract: 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.
authors.affiliation: Department of Basic Medical Sciences and Program in Molecular Cell Biology, Zhejiang University School of Medicine, Hangzhou 310058, China.
authors.forename: Mingjiao
authors.initials: M
authors.lastname: Sun
authors.forename: Huiying
authors.initials: H
authors.lastname: Fu
authors.forename: Hongqiang
authors.initials: H
authors.lastname: Cheng
authors.forename: Qian
authors.initials: Q
authors.lastname: Cao
authors.forename: Yuan
authors.initials: Y
authors.lastname: Zhao
authors.forename: Xiaozhou
authors.initials: X
authors.lastname: Mou
authors.forename: Xue
authors.initials: X
authors.lastname: Zhang
authors.forename: Xiuhua
authors.initials: X
authors.lastname: Liu
authors.forename: Yuehai
authors.initials: Y
authors.lastname: Ke
mesh_terms: D000199:Actins / Q000378:metabolism; D001671:Biochemistry / Q000379:methods*; D018938:Caco-2 Cells; D002463:Cell Membrane Permeability / Q000187:drug effects; D017097:Electric Impedance; D055664:Electrochemical Techniques; D000431:Ethanol / Q000633:toxicity; D006801:Humans; D007413:Intestinal Mucosa / Q000187:drug effects* / Q000378:metabolism; D052247:Nitric Oxide Synthase Type II / Q000037:antagonists & inhibitors / Q000378:metabolism; D047428:Protein Kinase Inhibitors / Q000494:pharmacology
chemical_list: D000199:Actins; D047428:Protein Kinase Inhibitors; D000431:Ethanol; D052247:Nitric Oxide Synthase Type II