title: Vulnerability of polarised intestinal porcine epithelial cells to mycotoxin deoxynivalenol depends on the route of application.
abstract: OBJECTIVE
  Deoxynivalenol (DON) is a Fusarium derived mycotoxin, often occurring on cereals used for human and animal nutrition. The intestine, as prominent barrier for nutritional toxins, has to handle the mycotoxin from the mucosa protected luminal side (apical exposure), as well as already absorbed toxin, reaching the cells from basolateral side via the blood stream. In the present study, the impact of the direction of DON exposure on epithelial cell behaviour and intestinal barrier integrity was elucidated.
  
  
  METHODS
  A non-transformed intestinal porcine epithelial cell line (IPEC-J2), cultured in membrane inserts, serving as a polarised in vitro model to determine the effects of deoxynivalenol (DON) on cellular viability and tight junction integrity.
  
  
  RESULTS
  Application of DON in concentrations up to 4000 ng/mL for 24, 48 and 72 hours on the basolateral side of membrane cultured polarised IPEC-J2 cells resulted in a breakdown of the integrity of cell connections measured by transepithelial electrical resistance (TEER), as well as a reduced expression of the tight junction proteins ZO-1 and claudin 3. Epithelial cell number decreased and nuclei size was enlarged after 72 h incubation of 4000 ng/mL DON from basolateral. Although necrosis or caspase 3 mediated apoptosis was not detectable after basolateral DON application, cell cycle analysis revealed a significant increase in DNA fragmentation, decrease in G0/G1 phase and slight increase in G2/M phase after 72 hours incubation with DON 2000 ng/mL.
  
  
  CONCLUSIONS
  Severity of impact of the mycotoxin deoxynivalenol on the intestinal epithelial barrier is dependent on route of application. The epithelium appears to be rather resistant towards apical (luminal) DON application whereas the same toxin dose from basolateral severely undermines barrier integrity.
authors.affiliation: Medical Faculty, Institute of Anatomy, Otto-von-Guericke University, Magdeburg, Germany.
authors.forename: Anne-Kathrin
authors.initials: AK
authors.lastname: Diesing
authors.forename: Constanze
authors.initials: C
authors.lastname: Nossol
authors.forename: Sven
authors.initials: S
authors.lastname: Dänicke
authors.forename: Nicole
authors.initials: N
authors.lastname: Walk
authors.forename: Andreas
authors.initials: A
authors.lastname: Post
authors.forename: Stefan
authors.initials: S
authors.lastname: Kahlert
authors.forename: Hermann-Josef
authors.initials: HJ
authors.lastname: Rothkötter
authors.forename: Jeannette
authors.initials: J
authors.lastname: Kluess
mesh_terms: D000818:Animals; D016764:Cell Polarity / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004333:Drug Administration Routes; D004351:Drug Resistance / Q000187:drug effects / Q000502:physiology; D017097:Electric Impedance; D004847:Epithelial Cells / Q000187:drug effects* / Q000502:physiology; D007413:Intestinal Mucosa / Q000166:cytology / Q000187:drug effects*; D009183:Mycotoxins / Q000494:pharmacology; D013552:Swine; D019108:Tight Junctions / Q000187:drug effects / Q000378:metabolism; D013997:Time Factors; D014255:Trichothecenes / Q000008:administration & dosage* / Q000494:pharmacology*
chemical_list: D009183:Mycotoxins; D014255:Trichothecenes; C007262:deoxynivalenol