Trolox and ascorbic acid reduce direct and indirect oxidative stress in the IPEC-J2 cells, an in vitro model for the porcine gastrointestinal tract.
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.
Laboratory of Applied Veterinary Morphology, Department of Veterinary Sciences, Faculty of Biomedical, Pharmaceutical and Veterinary Sciences, University of Antwerp, Wilrijk, Belgium.
Hans
H
Vergauwen
Laboratory of Applied Veterinary Morphology, Department of Veterinary Sciences, Faculty of Biomedical, Pharmaceutical and Veterinary Sciences, University of Antwerp, Wilrijk, Belgium.
Bart
B
Tambuyzer
Laboratory of Applied Veterinary Morphology, Department of Veterinary Sciences, Faculty of Biomedical, Pharmaceutical and Veterinary Sciences, University of Antwerp, Wilrijk, Belgium.
Karen
K
Jennes
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.
Jeroen
J
Degroote
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.
Wei
W
Wang
Laboratory for Animal Nutrition and Animal Product Quality (LANUPRO), Departement of Animal Production, Faculty of Bioscience Engineering, Ghent University, Melle, Belgium.
Stefaan
S
De Smet
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.
Joris
J
Michiels
Laboratory of Applied Veterinary Morphology, Department of Veterinary Sciences, Faculty of Biomedical, Pharmaceutical and Veterinary Sciences, University of Antwerp, Wilrijk, Belgium.
Chris
C
Van Ginneken
D000818:Animals; D000975:Antioxidants / Q000494:pharmacology*; D002460:Cell Line; D002839:Chromans / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D004341:Drug Evaluation; D041981:Gastrointestinal Tract / Q000166:cytology / Q000378:metabolism*; D008954:Models, Biological; D018384:Oxidative Stress / Q000187:drug effects*; D013552:Swine; D062826:Zonula Occludens-1 Protein / Q000378:metabolism
D000975:Antioxidants; D002839:Chromans; D062826:Zonula Occludens-1 Protein; C010643:6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid