In Vitro Investigation of Six Antioxidants for Pig Diets
Hans Vergauwen,
Sara Prims,
Jeroen Degroote,
Wei Wang,
Christophe Casteleyn,
Steven van Cruchten,
Stefaan de Smet,
Joris Michiels,
Chris van Ginneken
Affiliations
Hans Vergauwen
Laboratory of Applied Veterinary Morphology, Department of Veterinary Sciences, Faculty of Biomedical, University of Antwerp, Wilrijk 2610, Belgium
Sara Prims
Laboratory of Applied Veterinary Morphology, Department of Veterinary Sciences, Faculty of Biomedical, University of Antwerp, Wilrijk 2610, Belgium
Jeroen Degroote
Department of Applied Biosciences, Faculty of Bioscience Engineering, Ghent University, Ghent 9000, Belgium
Wei Wang
Department of Applied Biosciences, Faculty of Bioscience Engineering, Ghent University, Ghent 9000, Belgium
Christophe Casteleyn
Laboratory of Applied Veterinary Morphology, Department of Veterinary Sciences, Faculty of Biomedical, University of Antwerp, Wilrijk 2610, Belgium
Steven van Cruchten
Laboratory of Applied Veterinary Morphology, Department of Veterinary Sciences, Faculty of Biomedical, University of Antwerp, Wilrijk 2610, Belgium
Stefaan de Smet
Laboratory for Animal Nutrition and Animal Product Quality (LANUPRO), Department of Animal Production, Faculty of Bioscience Engineering, Ghent University, Melle 9090, Belgium
Joris Michiels
Department of Applied Biosciences, Faculty of Bioscience Engineering, Ghent University, Ghent 9000, Belgium
Chris van Ginneken
Laboratory of Applied Veterinary Morphology, Department of Veterinary Sciences, Faculty of Biomedical, University of Antwerp, Wilrijk 2610, Belgium
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.
