Journal Article

Fatty Acid Ethyl Esters Induce Intestinal Epithelial Barrier Dysfunction via a Reactive Oxygen Species-Dependent Mechanism in a Three-Dimensional Cell Culture Model

Troost, Freddy, Masclee, Ad, Jonkers, Daisy, Juuti-Uusitalo, Kati, van IJzendoorn, Sven, Elamin, Elhaseen, Pieters, Harm-Jan, Dekker, Jan

2013

Overview

Evidence is accumulating that ethanol and its oxidative metabolite, acetaldehyde, can disrupt intestinal epithelial integrity, an important factor contributing to ethanol-induced liver injury. However, ethanol can also be metabolized non-oxidatively generating phosphatidylethanol and fatty acid ethyl esters (FAEEs). This study aims to investigate the effects of FAEEs on barrier function, and to explore the role of oxidative stress as possible mechanism. Epithelial permeability was assessed by paracellular flux of fluorescein isothiocyanate-conjugated dextran using live cell imaging. Cell integrity was evaluated by lactate dehydrogenase release. Localization and protein levels of ZO-1 and occludin were analyzed by immunofluorescence and cell-based ELISA, respectively. Intracellular oxidative stress and cellular ATP levels were measured by dichlorofluorescein and luciferase driven bioluminescence, respectively. In vitro, ethyl oleate and ethyl palmitate dose dependently increased permeability associated with disruption and decreased ZO-1 and occludin protein levels, respectively, and increased intracellular oxidative stress without compromising cell viability. These effects could partially be attenuated by pretreatment with the antioxidant, resveratrol, pointing to the role of oxidative stress in the FAEEs-induced intestinal barrier dysfunction. These findings show that FAEEs can induce intestinal barrier dysfunction by disrupting the tight junctions, most likely via reactive oxygen species-dependent mechanism.

• Publisher: Public Library of Science,Public Library of Science (PLoS)
• Subject: Acetaldehyde / acetaldehyde-induced increase / Adenosine Triphosphate - metabolism / Antioxidants / Biology / Bioluminescence / Caco-2 Cells / Cell culture / Cell Culture Techniques / Cell Membrane Permeability - drug effects / Chemistry / Dextran / Dextrans / Enzyme-linked immunosorbent assay / Esters / Ethanol / Ethanol - metabolism / Ethanol - toxicity / Ethical aspects / Ethyl esters / Fatty acids / Fluorescein / Fluorescein isothiocyanate / heavy drinkers / Humans / hydrogen-peroxide / Immunofluorescence / Integrity / Intestinal Mucosa - drug effects / Intestinal Mucosa - physiopathology / Intestine / Intracellular / Isothiocyanate / kappa-b activation / Kinases / L-Lactate dehydrogenase / Lactate dehydrogenase / Lactic acid / Liver / Localization / Luciferase / Medicine / Metabolites / Models, Biological / nonoxidative metabolites / Nutrition / Occludin - metabolism / Oleic Acids - metabolism / Oleic Acids - toxicity / Oxidative Stress / Oxygen / Palmitic acid / Palmitic Acids - metabolism / Palmitic Acids - toxicity / pancreatic acinar-cells / paracellular permeability / Permeability / Pretreatment / Reactive oxygen species / Reactive Oxygen Species - metabolism / Resveratrol / Rodents / Small intestine / Stilbenes - pharmacology / Three dimensional models / tight junction proteins / Tight junctions / Tight Junctions - drug effects / Tight Junctions - metabolism / tyrosine phosphorylation / Zonula occludens-1 protein / Zonula Occludens-1 Protein - metabolism