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| Deoxynivalenol Induces Inflammation in IPEC-J2 Cells by ActivatingTXNIP/NLRP3 Inflammasome Pathway |
| ZHOU Chuang1,2, XIA Ying1, DING Shuo1, WANG Xing-Di1, WEI Xiao-Jun1, SHI Lu1, ZHANG Hua1* |
1 College of Animal Husbandry and Veterinary Medicine, Jiangsu Polytechnic College Agriculture and Forestry, Jurong 212400, China;
2 College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, China |
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Abstract Deoxynivalenol (DON) is one of the most serious mycotoxins which severely contaminates grains and their products. Exposure to DON causes animal poisoning, and its primary target organ is the intestine. To explore the connection and mechanisms involved in the intestinal inflammatory response triggered by DONand its interaction with the NOD-like receptor signaling pathway, in this study, porcine intestinal epithelial cells (IPEC-J2) were cultured with 1 μg/mL DON. ELISA was used to quantify the concentration of interleukin-1β (IL-1β) in the cell supernatants. The expression levels of thioredoxin interaction protein (TXNIP), cysteinyl aspartate specific proteinase-1 P20 (Caspase-1 P20) and NOD-like receptor protein 3 (NLRP3) were detected by Western blot. TXNIP siRNA transfection was used to further verify the regulation of NLRP3 by TXNIP. Antioxidant N-acetyl-cysteine (NAC) was used to detect the interaction among reactive oxygen species (ROS) and NLRP3 inflammasome activation. The results showed that compared with blank control group, the secretion of IL-1β in the cell supernatant was extremely significantly elevated in the DON-treated group. (P<0.01), and the protein expressions of TXNIP/NLRP3 inflammasome pathway-relatedproteins TXNIP, NLRP3 and Caspase-1 P20 in the cells were significantly up-regulated (P<0.05). Compared with negative transfection in DON-treated group, the secretion of IL-1β in the supernatant of TXNIP siRNA transfection in DON-treated group was extremely significantly decreased (P<0.01), and the proteinexpression of Caspase-1 P20 in the cells was significantly down-regulated (P<0.05). Compared with DON- treated group, the NAC intervention group exhibited significantly reduced cellular ROS levels (P<0.05), decreased secretion of IL-1β in the supernatant, and significantly downregulated expression of TXNIP andCaspase-1 P20 protein levels (P<0.05) in the cells. This study showed that DON induced the inflammatory response of IPEC-J2 cells through the TXNIP/NLRP3 inflammasome pathway, and the antioxidant NAC could alleviate the activation of NLRP3 induced by DON. This study identifies novel therapeutic targets and offers new insights for the effective prevention and management of DON-induced toxicity.
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Received: 11 June 2024
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Corresponding Authors:
*zhanghuasd@163.com
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