Deoxynivalenol and its metabolite deepoxy-deoxynivalenol: multi-parameter analysis for the evaluation of cytotoxicity and cellular effects

Abstract

The mycotoxin deoxynivalenol (DON) contaminates agricultural commodities worldwide, posing health threats to humans and animals. Associated with DON are derivatives, such as deepoxy-deoxynivalenol (DOM-1), produced by enzymatic transformation of certain intestinal bacteria, which are naturally occurring or applied as feed additives. Using differentiated porcine intestinal epithelial cells (IPEC-J2), we provide the first multi-parameter comparative cytotoxicity analysis of DON and DOM-1, based on the parallel evaluation of lysosomal activity, total protein content, membrane integrity, mitochondrial metabolism and ATP synthesis. The study investigated the ability of DON and-for the first time of its metabolite DOM-1-to induce apoptosis, mitogenactivated protein kinase (MAPK) signalling, oxidative events and alterations of mitochondrial structure in porcine intestinal epithelial cells (IECs). The degree of DON toxicity strongly varied, depending on the cytotoxicity parameter evaluated. DON compromised viability according to the parameters of lysosomal activity, total protein content and membrane integrity, but increased viability according to assays based on mitochondrial metabolism and ATP synthesis. DON induced expression of cleaved caspase-3 (maximum induction 3.9-fold) and MAPK p38 and p42/p44 (maximum induction 2.51-and 2.30-fold, respectively). DON altered mitochondrial morphology, but did not increase intracellular ROS. DOM-1-treated IPEC-J2 remained unaffected at equimolar concentrations in all assays, thereby confirming the safety of feed additives using DON-to DOM-1-transforming bacteria. The study additionally highlights that an extensive multi-parameter analysis significantly contributes to the quality of in vitro data.

Chronic ingestion of deoxynivalenol‐contaminated diet dose‐dependently decreases the area of myenteric neurons and gliocytes of rats

“…At the concentrations used in this work, the oxidative parameters analyzed were not changed. Springler et al measured the oxidative status in intestinal cell cultures exposed to DON and did not find increased ROS, although they did observe altered mitochondrial morphology. Investigations demonstrating that the mitochondrial changes caused by DON can alter the cell oxidative status were carried out using doses higher than those of this work.…”

Deoxynivalenol: Masked forms, fate during food processing, and potential biological remedies

“…In chickens and turkeys, DOM‐1 was metabolized as DOM‐3‐sulfate by oral ingestion (Schwartz‐Zimmermann et al., 2015). DOM‐1 did not activate the phosphorylation of the MAPK pathway in IPEC‐J2 cells, Caco‐2 cells, or jejunal explants, such as p38/MAPK, JNK/MAPK and ERK 1/2 /MAPK, and Sapk/JNK (Pierron, 2016; Springler et al., 2017). DOM‐1 retained the ability to fit into the A‐site of the ribosome peptidyl transferase, and only two hydrogen bonds were formed.…”

“…DOM‐1 showed no significant effects on the viability of RAW 264.7 cells, Jurkat T cells, RTgill‐W1, IPEC‐1, Caco‐2 cells, and IPEC‐J2. In addition, DOM‐1 hardly influenced membrane integrity, barrier function, cellular metabolism, intracellular ATP, caspase‐3, GSH/GSSG ratio, reactive oxygen species (Springler et al., 2017), mitochondrial activity, cytochrome c, and Bcl‐2 (Nasri, Bosch, Ten Voorde, & Fink‐Gremmels, 2006). The IC 50 of DOM‐1 was 55 times higher than that of DON in 3T3 fibroblasts (Eriksen, Pettersson, & Lundh, 2004).…”

Deoxynivalenol reduces quality parameters and increases DNA damage in mice spermatozoa

“…The given concentrations were selected based on previously published studies (Malekinejad et al, 2007;Pinton et al, 2012;Springler et al, 2017). The proposed concentrations of DON (0, 2.5, 5 and 10 μM) were prepared by using Ham's F10 culture medium (Sigma-Aldrich, Germany).…”