title: In Vitro Investigation of Six Antioxidants for Pig Diets.
abstract: Oxidative stress in the small intestinal epithelium can lead to barrier malfunction. In this study, the effect of rosmarinic acid (RA), quercetin (Que), gallic acid (GA), lipoic acid (LA), ethoxyquin (ETQ) and Se-methionine (SeMet) pre-treatments using 2 mM Trolox as a control on the viability and the generation of intracellular reactive oxygen species (iROS) of oxidatively (H₂O₂) stressed intestinal porcine epithelial cells (IPEC-J2) was investigated. A neutral red assay showed that RA (50-400 µM), Que (12.5-200 µM), GA (50-400 µM), ETQ (6.25-100 µM), and SeMet (125-1000 µM) pre-treatments but not LA significantly increased the viability of H₂O₂-stressed IPEC-J2 cells (p< 0.05). A 5-(and-6)-chloromethyl-2',7'-dichlorodihydrofluorescein diacetate, acetyl ester (CM-H₂DCFDA) fluorescent probe showed that RA (100-600 µM), Que (25-800 µM), ETQ (3.125-100 µM) and SeMet (500-2000 µM) pre-treatments significantly reduced iROS in IPEC-J2 monolayers (p< 0.05). Moreover, RA and Que were most effective in reducing iROS. Therefore, the effects of RA and Que on barrier functioning in vitro were examined. RA and Que pre-treatments significantly decreased fluorescein isothiocyanate (FITC)-conjugated dextran-4 (4 kDa) permeability and transepithelial electrical resistance (TEER) of an IPEC-J2 cell monolayer (p< 0.05). These in vitro results of RA and Que hold promise for their use as antioxidants in pig feed.
authors.affiliation: Laboratory of Applied Veterinary Morphology, Department of Veterinary Sciences, Faculty of Biomedical, University of Antwerp, Wilrijk 2610, Belgium. hans.vergauwen@uantwerpen.be.
authors.forename: Hans
authors.initials: H
authors.lastname: Vergauwen
authors.affiliation: Laboratory of Applied Veterinary Morphology, Department of Veterinary Sciences, Faculty of Biomedical, University of Antwerp, Wilrijk 2610, Belgium. sara.prims@uantwerpen.be.
authors.forename: Sara
authors.initials: S
authors.lastname: Prims
authors.affiliation: Department of Applied Biosciences, Faculty of Bioscience Engineering, Ghent University, Ghent 9000, Belgium. jerdgroo.degroote@ugent.be.
authors.forename: Jeroen
authors.initials: J
authors.lastname: Degroote
authors.affiliation: Department of Applied Biosciences, Faculty of Bioscience Engineering, Ghent University, Ghent 9000, Belgium. Wei.Wang@nusciencegroup.com.|Laboratory for Animal Nutrition and Animal Product Quality (LANUPRO), Department of Animal Production, Faculty of Bioscience Engineering, Ghent University, Melle 9090, Belgium. Wei.Wang@nusciencegroup.com.
authors.forename: Wei
authors.initials: W
authors.lastname: Wang
authors.affiliation: Laboratory of Applied Veterinary Morphology, Department of Veterinary Sciences, Faculty of Biomedical, University of Antwerp, Wilrijk 2610, Belgium. christophe.casteleyn@uantwerpen.be.
authors.forename: Christophe
authors.initials: C
authors.lastname: Casteleyn
authors.affiliation: Laboratory of Applied Veterinary Morphology, Department of Veterinary Sciences, Faculty of Biomedical, University of Antwerp, Wilrijk 2610, Belgium. steven.vancruchten@uantwerpen.be.
authors.forename: Steven
authors.initials: S
authors.lastname: van Cruchten
authors.affiliation: Laboratory for Animal Nutrition and Animal Product Quality (LANUPRO), Department of Animal Production, Faculty of Bioscience Engineering, Ghent University, Melle 9090, Belgium. stefaan.desmet@ugent.be.
authors.forename: Stefaan
authors.initials: S
authors.lastname: de Smet
authors.affiliation: Department of Applied Biosciences, Faculty of Bioscience Engineering, Ghent University, Ghent 9000, Belgium. joris.michiels@ugent.be.
authors.forename: Joris
authors.initials: J
authors.lastname: Michiels
authors.affiliation: Laboratory of Applied Veterinary Morphology, Department of Veterinary Sciences, Faculty of Biomedical, University of Antwerp, Wilrijk 2610, Belgium. chris.vanginneken@uantwerpen.be.
authors.forename: Chris
authors.initials: C
authors.lastname: van Ginneken
keywords: IPEC-J2 cells; antioxidant pre-treatment; hydrogen peroxide; oxidative stress; small intestinal epithelial permeability