Zearalenone-Induced Mechanical Damage of Intestinal Barrier Via the RhoA/ROCK Signaling Pathway in IPEC-J2 Cells

Overview

Journal Int J Mol Sci

Publisher MDPI

Specialties Biochemistry
Chemistry
Molecular Biology

Date 2022 Oct 27

PMID 36293400

Authors

Biying Huang

Jingjing Wang

Aixin Gu

Tianhu Wang

Jianping Li

Anshan Shan

Affiliations

Soon will be listed here.

Abstract

Zearalenone (ZEN) is a widespread contaminant of cereals and agricultural products which causes food safety issues. Ingesting food or feed contaminated with ZEN can disrupt the intestinal epithelial barrier function. The RhoA/ROCK signaling pathway plays a key role in regulating the epithelial barrier function, but studies on such roles have rarely focused on the intestine. The aim of this experiment was to investigate the exact mechanism of ZEN-induced intestinal barrier damage and whether the RhoA/ROCK signaling pathway is involved. The results showed that ZEN significantly induced alkaline phosphatase (AP) activity and FITC-dextran (4 kDa) passage across the epithelial barrier, which significantly reduced the transepithelial resistance (TEER). Meanwhile, ZEN could induce the significantly down-regulated mRNA expression of tight junction proteins (occludin, claudin-1, ZO-1, and claudin-3) and redistribution of ZO-1 immunofluorescence. Further studies demonstrated that ZEN exposure activated the RhoA/ROCK signaling pathway, significantly up-regulated the mRNA expression of ROCK1, the main effector of the signaling pathway, the protein expression of phosphorylated myosin light chain (MLC) and myosin light chain kinase (MLCK), and relatively increased the activity of ATP in cells, simultaneously remodeling the cytoskeleton (F-actin). Overall, our study indicated that ZEN induced intestinal barrier dysfunction by activating the RhoA/ROCK signaling pathway.

Citing Articles

Butyric acid alleviates LPS-induced intestinal mucosal barrier damage by inhibiting the RhoA/ROCK2/MLCK signaling pathway in Caco2 cells.

Liu L, Chen T, Xie Z, Zhang Y, He C, Huang Y PLoS One. 2024; 19(12):e0316362.

PMID: 39724098 PMC: 11670954. DOI: 10.1371/journal.pone.0316362.

Asiaticoside ameliorates uterine injury induced by zearalenone in mice by reversing endometrial barrier disruption, oxidative stress and apoptosis.

Gao G, Jiang H, Lin H, Yang H, Wang K Reprod Biol Endocrinol. 2024; 22(1):118.

PMID: 39272165 PMC: 11395188. DOI: 10.1186/s12958-024-01288-6.

Positive effects of selenized-oligochitosan on zearalenone-induced intestinal dysfunction in piglets.

Qin S, Peng Y, She F, Zhang J, Li L, Chen F Front Vet Sci. 2023; 10:1184969.

PMID: 37261113 PMC: 10228365. DOI: 10.3389/fvets.2023.1184969.

Effects of Exposure to Low Zearalenone Concentrations Close to the EU Recommended Value on Weaned Piglets' Colon.

Grosu I, Bulgaru C, Pistol G, Cismileanu A, Marin D, Taranu I Toxins (Basel). 2023; 15(3).

PMID: 36977097 PMC: 10055674. DOI: 10.3390/toxins15030206.

References

Fan W, Shen T, Ding Q, Lv Y, Li L, Huang K . Zearalenone induces ROS-mediated mitochondrial damage in porcine IPEC-J2 cells. J Biochem Mol Toxicol. 2017; 31(10). DOI: 10.1002/jbt.21944. View

Pastorelli L, De Salvo C, Mercado J, Vecchi M, Pizarro T . Central role of the gut epithelial barrier in the pathogenesis of chronic intestinal inflammation: lessons learned from animal models and human genetics. Front Immunol. 2013; 4:280. PMC: 3775315. DOI: 10.3389/fimmu.2013.00280. View

Utech M, Ivanov A, Samarin S, Bruewer M, Turner J, Mrsny R . Mechanism of IFN-gamma-induced endocytosis of tight junction proteins: myosin II-dependent vacuolarization of the apical plasma membrane. Mol Biol Cell. 2005; 16(10):5040-52. PMC: 1237102. DOI: 10.1091/mbc.e05-03-0193. View

Devreese M, Pasmans F, De Backer P, Croubels S . An in vitro model using the IPEC-J2 cell line for efficacy and drug interaction testing of mycotoxin detoxifying agents. Toxicol In Vitro. 2012; 27(1):157-63. DOI: 10.1016/j.tiv.2012.09.020. View

Liao P, Liao M, Li L, Tan B, Yin Y . Effect of deoxynivalenol on apoptosis, barrier function, and expression levels of genes involved in nutrient transport, mitochondrial biogenesis and function in IPEC-J2 cells. Toxicol Res (Camb). 2018; 6(6):866-877. PMC: 6060741. DOI: 10.1039/c7tx00202e. View

Horowitz S, Binion D, Nelson V, Kanaa Y, Javadi P, Lazarova Z . Increased arginase activity and endothelial dysfunction in human inflammatory bowel disease. Am J Physiol Gastrointest Liver Physiol. 2007; 292(5):G1323-36. DOI: 10.1152/ajpgi.00499.2006. View

Dong N, Xue C, Zhang L, Zhang T, Wang C, Bi C . Oleanolic acid enhances tight junctions and ameliorates inflammation in Salmonella typhimurium-induced diarrhea in mice via the TLR4/NF-κB and MAPK pathway. Food Funct. 2019; 11(1):1122-1132. DOI: 10.1039/c9fo01718f. View

Tong J, Wang Y, Chang B, Zhang D, Wang B . Y-27632 inhibits ethanol-induced increase in intestinal epithelial barrier permeability. Mol Med Rep. 2014; 9(6):2357-61. DOI: 10.3892/mmr.2014.2060. View

Guo W, Wang P, Liu Z, Ye P . Analysis of differential expression of tight junction proteins in cultured oral epithelial cells altered by Porphyromonas gingivalis, Porphyromonas gingivalis lipopolysaccharide, and extracellular adenosine triphosphate. Int J Oral Sci. 2018; 10(1):e8. PMC: 5795020. DOI: 10.1038/ijos.2017.51. View

10.

Tralamazza S, Piacentini K, Savi G, Carnielli-Queiroz L, de Carvalho Fontes L, Martins C . Wild rice (O. latifolia) from natural ecosystems in the Pantanal region of Brazil: Host to Fusarium incarnatum-equiseti species complex and highly contaminated by zearalenone. Int J Food Microbiol. 2021; 345:109127. DOI: 10.1016/j.ijfoodmicro.2021.109127. View

11.

Wang J, Li M, Zhang W, Gu A, Dong J, Li J . Protective Effect of N-Acetylcysteine against Oxidative Stress Induced by Zearalenone via Mitochondrial Apoptosis Pathway in SIEC02 Cells. Toxins (Basel). 2018; 10(10). PMC: 6215273. DOI: 10.3390/toxins10100407. View

12.

Gil-Cardoso K, Gines I, Pinent M, Ardevol A, Blay M, Terra X . Effects of flavonoids on intestinal inflammation, barrier integrity and changes in gut microbiota during diet-induced obesity. Nutr Res Rev. 2016; 29(2):234-248. DOI: 10.1017/S0954422416000159. View

13.

Sun L, Lei M, Zhang N, Zhao L, Krumm C, Qi D . Hepatotoxic effects of mycotoxin combinations in mice. Food Chem Toxicol. 2014; 74:289-93. DOI: 10.1016/j.fct.2014.10.020. View

14.

Putt K, Pei R, White H, Bolling B . Yogurt inhibits intestinal barrier dysfunction in Caco-2 cells by increasing tight junctions. Food Funct. 2017; 8(1):406-414. DOI: 10.1039/c6fo01592a. View

15.

Sun X, Shi J, Zou X, Wang C, Yang Y, Zhang H . Silver nanoparticles interact with the cell membrane and increase endothelial permeability by promoting VE-cadherin internalization. J Hazard Mater. 2016; 317:570-578. DOI: 10.1016/j.jhazmat.2016.06.023. View

16.

Zhou H, Zhu H, Yao X, Qian J, Yang J, Pan X . Metformin regulates tight junction of intestinal epithelial cells via MLCK-MLC signaling pathway. Eur Rev Med Pharmacol Sci. 2017; 21(22):5239-5246. DOI: 10.26355/eurrev_201711_13847. View

17.

Tasaka S, Koh H, Yamada W, Shimizu M, Ogawa Y, Hasegawa N . Attenuation of endotoxin-induced acute lung injury by the Rho-associated kinase inhibitor, Y-27632. Am J Respir Cell Mol Biol. 2005; 32(6):504-10. DOI: 10.1165/rcmb.2004-0009OC. View

18.

Pinton P, Nougayrede J, Del Rio J, Moreno C, Marin D, Ferrier L . The food contaminant deoxynivalenol, decreases intestinal barrier permeability and reduces claudin expression. Toxicol Appl Pharmacol. 2009; 237(1):41-8. DOI: 10.1016/j.taap.2009.03.003. View

19.

Odenwald M, Choi W, Buckley A, Shashikanth N, Joseph N, Wang Y . ZO-1 interactions with F-actin and occludin direct epithelial polarization and single lumen specification in 3D culture. J Cell Sci. 2016; 130(1):243-259. PMC: 5394778. DOI: 10.1242/jcs.188185. View

20.

Xie H, Xue Y, Liu L, Liu Y . Endothelial-monocyte-activating polypeptide II increases blood-tumor barrier permeability by down-regulating the expression levels of tight junction associated proteins. Brain Res. 2010; 1319:13-20. DOI: 10.1016/j.brainres.2010.01.023. View