Deoxynivalenol Induces Inflammatory Injury in IPEC-J2 Cells via NF-κB Signaling Pathway

Abstract

The aim of this study was to investigate the effects of deoxynivalenol (DON) exposure on the inflammatory injury nuclear factor kappa-B (NF-κB) pathway in intestinal epithelial cells (IPEC-J2 cells) of pig. The different concentrations of DON (0, 125, 250, 500, 1000, 2000 ng/mL) were added to the culture solution for treatment. The NF-κB pathway inhibitor pyrrolidine dithiocarbamate (PDTC) was used as a reference. The results showed that when the DON concentration increased, the cell density decreased and seemed damaged. With the increase of DON concentration in the culture medium, the action of diamine oxidase (DAO) in the culture supernatant also increased. The activities of IL-6, TNF-α, and NO in the cells were increased with the increasing DON concentration. The relative mRNA expression of IL-1β and IL-6 were increased in the cells. The mRNA relative expression of NF-κB p65, IKKα, and IKKβ were upregulated with the increasing of DON concentration, while the relative expression of IκB-α mRNA was downregulated. At the same time, the expression of NF-κB p65 protein increased gradually in the cytoplasm and nucleus with a higher concentration of DON. These results showed that DON could change the morphology of IPEC-J2 cells, destroy its submicroscopic structure, and enhance the permeability of cell membrane, as well as upregulate the transcription of some inflammatory factors and change the expression of NF-κB-related gene or protein in cells.

In-Vitro Cell Culture for Efficient Assessment of Mycotoxin Exposure, Toxicity and Risk Mitigation

“…At 2 µM of DON exposure, DON has been reported to up-regulate the expression of IPEC-J2 cell IL1-α, IL1-β, IL-6, IL-8, TNF-α, and MCP1 genes after 48 h of exposure, whereas, 0.5 µM exposure simulated expression of IL1-β, IL-6, and IL-8 genes, and down-regulated expression of IL1-α and MCP1 genes [200]. The up-regulatory effect of DON on IL-6, TNF-α, and IL1-β through the NF-κB pathway was also observed in IPEC-J2 cells after 24 h of DON exposure within the concentration range of 0.34 µM to 6.7 µM [162]. DON also reportedly induced a concentration-dependent increase in the secretion of IL-8 protein by Caco-2 cells through NF-κB after 48 h exposure [201], and [194] reported an increase in IL-8 protein secretion by Caco-2 cells after 12 h, which was associated with NF-κB, PKR, and p38 pathways.…”

Nontoxic-dose deoxynivalenol aggravates lipopolysaccharides-induced inflammation and tight junction disorder in IPEC-J2 cells through activation of NF-κB and LC3B

“…Next, we further studied the mechanism of the above phenomenon that nontoxic-dose DON could aggravated LPS-induced intestinal inflammation and barrier dysfunction in IPEC-J2 cell monolayers. Many studies have reported that mycotoxins could regulate inflammatory cytokines through regulating nuclear factor-κB (NF-κB) signaling pathway ( Shao et al, 2019 ; Sugiyama et al, 2016 ; Wang et al, 2019b ; Zmora et al, 2017 ). In addition, NF-κB has been reported to regulate the tight junction barrier and NLRP3 inflammasome response to some extracellular stimuli ( Al-Sadi et al, 2010 ; Guo et al, 2019; Zmora et al, 2017 ).…”

Transcriptome-wide assessment of the m6A methylome of intestinal porcine epithelial cells treated with deoxynivalenol

“…Wang et al (2018) revealed that DON treatment induces toxicity and apoptotic cell death in piglet hippocampal nerve cells via the MAPK signaling pathway [40]. Further, Zhang et al (2020) reported that in IPEC-J2 cells, DON treatment induced the production of proin ammatory factors by activating p38 and ERK1/2 [18], while showed that treating IPEC-J2 cells with DON caused in ammatory injury via activation of the NF-κB signaling pathway [41]. With regard to the m 6 A-modi ed genes identi ed in this study, our RNA-seq and qPCR analyses indicated that CSF2 was differentially expressed the most signi cantly in DON-treated cells, and CSF2 was the most central protein in the constructed PPI network (Fig.…”