title: Deoxynivalenol, but not E. coli lipopolysaccharide, changes the response pattern of intestinal porcine epithelial cells (IPEC-J2) according to its route of application.
abstract: The porcine intestinal epithelium is a primary target for mycotoxin deoxynivalenol (DON) and lipopolysaccharides (LPS). Although epithelial cells are exposed to these toxins mainly from the luminal-chyme compartment an exposure from the blood side resulting from systemic absorption cannot be excluded. Thus, we investigated the effect of DON and LPS, alone or combined, on porcine intestinal epithelial cells IPEC-J2 on a transcriptional, translational and functional level when administered either from apical or basolateral. IPEC-J2 cells were cultured on 12-well inserts in complete medium at 5% CO2 and 39°C and subjected to following treatments: control (CON), 2000 ng/mL DON, 1 μg/mL LPS or DON+LPS for 72 h, either from apical or basolateral. Transepithelial electrical resistance (TEER), protein and IL-8 content were measured and microarray analysis, qRT-PCR (IL-8, zonula occludens-1 ZO-1, β-actin), Western Blot (ZO-1, β-actin) and immunofluorescence (ZO-1) were performed. Data of at least three independent experiments were analysed with ANOVA and Dunnett's post hoc test. Basolateral DON resulted in significantly lower cell counts (p<0.05) with larger cells (p<0.01), whereas apical DON reduced total (p<0.001) and specific protein content (IL-8 content CON vs. DON: 2378 pg/3 mL vs. 991 pg/3 mL; p<0.001). Transcripts of ß-actin and ZO-1 were significantly upregulated in response to DON, irrespective of direction, whereas IL-8 mRNA remained unaffected. However, ZO-1 spatial distribution in the tight junction and its function (TEER) were detrimentally affected by basolateral DON only. In conclusion, direction of DON exposure affected IPEC-J2 differently on a translational and functional level, but was mainly inconsequential on a transcriptional level.
authors.affiliation: Institute of Animal Nutrition, Friedrich-Loeffler-Institute, Federal Research Institute for Animal Health, Braunschweig, Germany. Electronic address: jeannette.kluess@fli.bund.de.
authors.forename: J W
authors.initials: JW
authors.lastname: Kluess
authors.affiliation: Institute of Anatomy, Otto von Guericke University Magdeburg, Magdeburg, Germany.
authors.forename: S
authors.initials: S
authors.lastname: Kahlert
authors.affiliation: Institute of Anatomy, Otto von Guericke University Magdeburg, Magdeburg, Germany.
authors.forename: A
authors.initials: A
authors.lastname: Kröber
authors.affiliation: Institute of Anatomy, Otto von Guericke University Magdeburg, Magdeburg, Germany.
authors.forename: A-K
authors.initials: AK
authors.lastname: Diesing
authors.affiliation: Institute of Anatomy, Otto von Guericke University Magdeburg, Magdeburg, Germany.
authors.forename: H-J
authors.initials: HJ
authors.lastname: Rothkötter
authors.affiliation: Genome Biology, Leibniz Institute for Farm Animal Biology, Dummerstorf, Germany.
authors.forename: Klaus
authors.initials: K
authors.lastname: Wimmers
authors.affiliation: Institute of Animal Nutrition, Friedrich-Loeffler-Institute, Federal Research Institute for Animal Health, Braunschweig, Germany.
authors.forename: S
authors.initials: S
authors.lastname: Dänicke
mesh_terms: D000199:Actins / Q000235:genetics / Q000378:metabolism; D000818:Animals; D002452:Cell Count; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000187:drug effects*; D004926:Escherichia coli / Q000737:chemistry; D016209:Interleukin-8 / Q000235:genetics / Q000378:metabolism; D007422:Intestines / Q000166:cytology / Q000187:drug effects; D008070:Lipopolysaccharides / Q000633:toxicity*; D013552:Swine; D019108:Tight Junctions / Q000187:drug effects / Q000378:metabolism; D014255:Trichothecenes / Q000633:toxicity*; D015854:Up-Regulation; D062826:Zonula Occludens-1 Protein / Q000235:genetics / Q000378:metabolism
chemical_list: D000199:Actins; D016209:Interleukin-8; D008070:Lipopolysaccharides; D014255:Trichothecenes; D062826:Zonula Occludens-1 Protein; C007262:deoxynivalenol
keywords: Deoxynivalenol; IL-8; IPEC-J2; Lipopolysaccharide; ZO-1