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Home » Archive » 2020

TDK conference 2020

Investigation of the beneficial effect of rosemary acid on deoxynivalenol and T-2 mycotoxin poisoning on IPEC-J2 cells
Szóládi Áron - year 6
University of Veterinary Medicine Budapest, Department of Pharmacology and Toxicology
Supervisor: Erzsébet dr. Pásztiné dr. Gere

Abstract:

Large-scale, intensive animal husbandry has a significant demand for grain. Due to global climate change, cereals cannot be harvested at the right time, increasing the chances of field and warehouse mold settling. Fusariotoxins are products of secondary metabolism in Fusarium species that have a detrimental effect on the gastrointestinal tract of the swine on contaminated corn-based feed leading to the development of retarded growth, weight loss, fertility problems and lowered productivity.

In our study, we investigated the cellular effects of deoxynivalenol (DON), T-2 and their combination among the fusarium toxins, as these toxins cause pronounced economic damage, as they cause feed rejection in farm animals, thus reducing the productivity of the animals. These mycotoxins generate excessive oxidative stress, which weakens the intestinal barrier integrity in swine.

In our experiments, we sought to answer how treatment with fusariotoxins affects the viability of porcine small intestinal IPEC-J2 cells, and which changes in the levels of inflammatory cytokines, in cell redox status and in intestinal barrier integrity can be detected, and how rosmarinic acid (RA) with antioxidant properties can alleviate these harmful effects.

The Neutral red measurement method was used to monitor changes in cell viability. Transepithelial resistance (TER) measurement was used to determine cell layer integrity, and the localization of claudin-1 and occludin in the tight junction was examined before and after treatment. Extracellular H2O2 levels were measured by the Amplex Red method. Inflammatory cytokine concentrations of interleukin (IL) -6 and IL-8 were determined by enzyme-linked immunosorbent assays (ELISA).

It was ascertained that simultaneous treatment of DON and T-2 (DT2: 1 µmol/L DON + 5 nmol/L T-2) for 48 h and 72 h reduced TER of the cell monolayer, which was restored by 50 µmol/L RA application. It was also found that DT2 for 48 and 72 h could induce oxidative stress and elevate IL-8 levels significantly, which was alleviated by administration of RA. DT2 administration contributed to redistribution of claudin-1 and triggered its loss from cell membrane; however, occludin membranous localization was not altered by combined mycotoxin treatment.

Based on our results, it can be predicted that in the case of confirmatory data from in vivo future experiments, supplementing the feed with RA can effectively reduce and compensate for the toxic intestinal effects in swine during the feeding of mycotoxin-contaminated grain.



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