Trolox and ascorbic acid reduce direct and indirect oxidative stress in the IPEC-J2 cells, an in vitro model for the porcine gastrointestinal tract

Hans Vergauwen; Bart Tambuyzer; Karen Jennes; Jeroen Degroote; Wei Wang; Stefaan De Smet; Joris Michiels; Chris Van Ginneken

doi:10.1371/journal.pone.0120485

Trolox and ascorbic acid reduce direct and indirect oxidative stress in the IPEC-J2 cells, an in vitro model for the porcine gastrointestinal tract

PLoS One. 2015 Mar 6;10(3):e0120485. doi: 10.1371/journal.pone.0120485. eCollection 2015.

Authors

Hans Vergauwen¹, Bart Tambuyzer¹, Karen Jennes¹, Jeroen Degroote², Wei Wang², Stefaan De Smet³, Joris Michiels², Chris Van Ginneken¹

Affiliations

¹ Laboratory of Applied Veterinary Morphology, Department of Veterinary Sciences, Faculty of Biomedical, Pharmaceutical and Veterinary Sciences, University of Antwerp, Wilrijk, Belgium.
² Department of Applied Biosciences, Faculty of Bioscience Engineering, Ghent University, Ghent, Belgium; Laboratory for Animal Nutrition and Animal Product Quality (LANUPRO), Departement of Animal Production, Faculty of Bioscience Engineering, Ghent University, Melle, Belgium.
³ Laboratory for Animal Nutrition and Animal Product Quality (LANUPRO), Departement of Animal Production, Faculty of Bioscience Engineering, Ghent University, Melle, Belgium.

Abstract

Oxidative stress in the small intestinal epithelium is a major cause of barrier malfunction and failure to regenerate. This study presents a functional in vitro model using the porcine small intestinal epithelial cell line IPEC-J2 to examine the effects of oxidative stress and to estimate the antioxidant and regenerative potential of Trolox, ascorbic acid and glutathione monoethyl ester. Hydrogen peroxide and diethyl maleate affected the tight junction (zona occludens-1) distribution, significantly increased intracellular oxidative stress (CM-H2DCFDA) and decreased the monolayer integrity (transepithelial electrical resistance and FD-4 permeability), viability (neutral red) and wound healing capacity (scratch assay). Trolox (2 mM) and 1 mM ascorbic acid pre-treatment significantly reduced intracellular oxidative stress, increased wound healing capacity and reduced FD-4 permeability in oxidatively stressed IPEC-J2 cell monolayers. All antioxidant pre-treatments increased transepithelial electrical resistance and viability only in diethyl maleate-treated cells. Glutathione monoethyl ester (10 mM) pre-treatment significantly decreased intracellular oxidative stress and monolayer permeability only in diethyl maleate-treated cells. These data demonstrate that the IPEC-J2 oxidative stress model is a valuable tool to screen antioxidants before validation in piglets.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Antioxidants / pharmacology*
Cell Line
Chromans / pharmacology*
Dose-Response Relationship, Drug
Drug Evaluation
Gastrointestinal Tract / cytology
Gastrointestinal Tract / metabolism*
Models, Biological*
Oxidative Stress / drug effects*
Swine
Zonula Occludens-1 Protein / metabolism

Substances

Antioxidants
Chromans
Zonula Occludens-1 Protein
6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid

Grants and funding

This work was supported by the IWT_LO 100856 project provided by the agency for Innovation by Science and Technology. (http://www.iwt.be/) (CVG SDS JM). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.