title: Mechanism and consequence of chitosan-mediated reversible epithelial tight junction opening.
abstract: In order to increase the absorption of hydrophilic macromolecules in the small intestine, permeation enhancers such as chitosan (CS) and its derivatives have been evaluated. The aim of the current work was to investigate, on molecular levels, the effect of CS on tight junction (TJ) integrity in Caco-2 cells. The observed changes in transepithelial-electrical-resistance measurements and the staining patterns of the monolayer Caco-2 cells demonstrate that CS can transiently and reversibly open the TJs between cells, thus enhancing the paracellular permeability. TJ ultra-structures examined by transmission electron microscopy support the concept that CS did induce transient opening of TJs. We then assessed TJ disruption at the gene and protein expression levels. Our data indicate that exposure to CS followed by recovery resulted in a significant increase in claudin-4 (Cldn4) gene transcription. Additionally, CS treatment induced redistribution of the TJ protein CLDN4 intracellularly following by its degradation in lysosomes, which represented an important contributing factor in TJ weakening, leading to the opening of TJs. The recovery of TJ after CS disruption required CLDN4 protein synthesis. These results suggest that CS regulates TJs by inducing changes in transmembrane CLDN4 protein. Understanding the mechanism of interaction between CS and epithelial cells is of paramount importance and needs to be established to aid further development in the use of CS to mediate the trans-epithelial drug delivery.
authors.affiliation: Department of Chemical Engineering, National Tsing Hua University, Hsinchu 30013, Taiwan, ROC.
authors.forename: Tzyy-Harn
authors.initials: TH
authors.lastname: Yeh
authors.forename: Li-Wen
authors.initials: LW
authors.lastname: Hsu
authors.forename: Michael T
authors.initials: MT
authors.lastname: Tseng
authors.forename: Pei-Ling
authors.initials: PL
authors.lastname: Lee
authors.forename: Kiran
authors.initials: K
authors.lastname: Sonjae
authors.forename: Yi-Cheng
authors.initials: YC
authors.lastname: Ho
authors.forename: Hsing-Wen
authors.initials: HW
authors.lastname: Sung
mesh_terms: D015153:Blotting, Western; D018938:Caco-2 Cells; D048271:Chitosan / Q000494:pharmacology; D062506:Claudin-4; D057167:Claudins / Q000378:metabolism; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D046529:Microscopy, Electron, Transmission; D016133:Polymerase Chain Reaction; D019108:Tight Junctions / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure
chemical_list: C568157:CLDN4 protein, human; D062506:Claudin-4; D057167:Claudins; D048271:Chitosan