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Zearalenone regulates key factors of the Kelch-like erythroid cell-derived protein with CNC homology-associated protein 1-nuclear factor erythroid 2-related factor 2 signaling pathway in duodenum of post-weaning gilts

  • Cheng, Qun (Shandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and Prevention, Department of Animal Sciences and Technology, Shandong Agricultural University) ;
  • Jiang, Shu zhen (Shandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and Prevention, Department of Animal Sciences and Technology, Shandong Agricultural University) ;
  • Huang, Li bo (Shandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and Prevention, Department of Animal Sciences and Technology, Shandong Agricultural University) ;
  • Yang, Wei ren (Shandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and Prevention, Department of Animal Sciences and Technology, Shandong Agricultural University) ;
  • Yang, Zai bin (Shandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and Prevention, Department of Animal Sciences and Technology, Shandong Agricultural University)
  • Received : 2020.06.02
  • Accepted : 2020.09.25
  • Published : 2021.08.01

Abstract

Objective: This study explored the mechanism of the Kelch-like erythroid cell-derived protein with CNC homology-associated protein 1 (Keap1)-nuclear factor erythroid 2-related factor 2 (Nrf2) signaling pathway under conditions of zearalenone (ZEA)-induced oxidative stress in the duodenum of post-weaning gilts. Methods: Forty post-weaning gilts were randomly allocated to four groups and fed diets supplemented with 0, 0.5, 1.0, or 1.5 mg/kg ZEA. Results: The results showed significant reductions in the activity of the antioxidant enzymes total superoxide dismutase and glutathione peroxidase and increases the malondialdehyde content with increasing concentrations of dietary ZEA. Immunohistochemical analysis supported these findings by showing a significantly increased expression of Nrf2 and glutathione peroxidase 1 (GPX1) with increasing concentrations of ZEA. The relative mRNA and protein expression of Nrf2, GPX1 increased linearly (p<0.05) and quadratically (p<0.05), which was consistent with the immunohistochemical results. The relative mRNA expression of Keap1 decreased linearly (p<0.05) and quadratically (p<0.05) in the duodenum as the ZEA concentration increased in the diet. The relative mRNA expression of modifier subunit of glutamate-cysteine ligase (GCLM) increased quadratically (p<0.05) in all ZEA treatment groups and the relative mRNA expression of quinone oxidoreductase 1 (NQO1) catalytic subunit of glutamate-cysteine ligase decreased linearly (p<0.05) and quadratically (p<0.05) in the ZEA1.0 group and ZEA1.5 group. The relative protein expression of Keap1 and GCLM decreased quadratically (p<0.05) in the duodenum as the ZEA concentration increased in the diet, respectively. The relative protein expression of NQO1 increased linearly (p<0.05) and quadratically (p<0.05) in all ZEA treatment groups in the duodenum. Conclusion: These findings suggest that ZEA regulates the expression of key factors of the Keap1-Nrf2 signaling pathway in the duodenum, which enables resistance to ZEA-induced oxidative stress. Further studies are needed to examine the effects of ZEA induced oxidative stress on other tissues and organs in post-weaning gilts.

Keywords

Acknowledgement

This study was funded in part by Major Innovative Projects in Shandong Province of Research and application of environment-friendly feed and the critical technologies for pigs and poultry without antibiotic (2019JZZY020609), the Natural Science Foundation of Shandong Province (Project No. ZR2019MC038), the Agriculture Research System in Shandong Province (Project No. SDAIT-08-05), and the Funds of Shandong "Double Tops".

References

  1. De Boevre M, Di Mavungu JD, Landschoot S, et al. Natural occurrence of mycotoxins and their masked forms in food and feed products. World Mycotoxin J 2012;5:207-19. https://doi.org/10.3920/WMJ2012.1410
  2. Chang H, Kim W, Park JH, et al. The occurrence of zearalenone in South Korean feedstuffs between 2009 and 2016. Toxins 2017;9:223. https://doi.org/10.3390/toxins9070223
  3. Price WD, Lovell RA, McChesney DG. Naturally occurring toxins in feedstuffs: center for veterinary medicine perspective. J Anim Sci 1993;71:2556-62. https://doi.org/10.2527/1993.7192556x
  4. Zinedine A, Soriano JM, Molto JC, Manes J. Review on the toxicity, occurrence, metabolism, detoxification, regulations and intake of zearalenone: an oestrogenic mycotoxin. Food Chem Toxicol 2007;45:1-18. https://doi.org/10.1016/j.fct.2006.07.030
  5. Takemura H, Shim JY, Sayama K, Tsubura A, Zhu BT, Shimoi K. Characterization of the estrogenic activities of zearalenone and zeranol in vivo and in vitro. J Steroid Biochem Mol Biol 2007;103:170-7. https://doi.org/10.1016/j.jsbmb.2006.08.008
  6. Wielogorska E, Elliott CT, Danaher M, Connolly L. Validation and application of a reporter gene assay for the determination of estrogenic endocrine disruptor activity in milk. Food Chem Toxicol 2014;69:260-6. https://doi.org/10.1016/j.fct.2014.04.028
  7. Dai M, Jiang S, Yuan X, Yang W, Yang Z, Huang L. Effects of zearalenone-diet on expression of ghrelin and PCNA genes in ovaries of post-weaning piglets. Anim Reprod Sci 2016; 168:126-37. https://doi.org/10.1016/j.anireprosci.2016.03.006
  8. Chen XX, Yang CW, Huang LB, Niu QS, Jiang SZ, Chi F. Zearalenone altered the serum hormones, morphologic and apoptotic measurements of genital organs in post-weaning gilts. Asian-Australas J Anim Sci 2015;28:171-9. https://doi.org/10.5713/ajas.14.0329
  9. Yang LJ, Zhou M, Huang LB, et al. Zearalenone-promoted follicle growth through modulation of Wnt-1/β-catenin signaling pathway and expression of estrogen receptor genes in ovaries of postweaning piglets. J Agric Food Chem 2018; 66:7899-906. https://doi.org/10.1021/acs.jafc.8b02101
  10. Reddy KE, Kim M, Kim KH, Ji SY, Lee SD. Effect of commercially purified deoxynivalenol and zearalenone mycotoxins on microbial diversity of pig cecum contents. Anim Biosci 2021;34:243-55. https://doi.org/10.5713/ajas.20.0137
  11. Li Y, Zhang B, Huang K, et al. Mitochondrial proteomic analysis reveals the molecular mechanisms underlying reproductive toxicity of zearalenone in MLTC-1 cells. Toxicology 2014; 324:55-67. https://doi.org/10.1016/j.tox.2014.07.007
  12. Golli-Bennour EE, Bacha H. Hsp70 expression as biomarkers of oxidative stress: mycotoxins' exploration. Toxicology 2011; 287:1-7. https://doi.org/10.1016/j.tox.2011.06.002
  13. Gajecka M, Rybarczyk L, Jakimiuk E, et al. The effect of experimental long-term exposure to low-dose zearalenone on uterine histology in sexually immature gilts. Exp Toxicol Pathol 2012; 64:537-42. https://doi.org/10.1016/j.etp.2010.11.009
  14. Schoevers EJ, Santos RR, Colenbrander B, Fink-Gremmels J, Roelen BAJ. Transgenerational toxicity of zearalenone in pigs. Reprod Toxicol 2012;34:110-9. https://doi.org/10.1016/j.reprotox.2012.03.004
  15. Biehl ML, Prelusky DB, Koritz GD, Hartin KE, Buck WB, Trenholm HL. Biliary excretion and enterohepatic cycling of zearalenone in immature pigs. Toxicol Appl Pharmacol 1993;121:152-9. https://doi.org/10.1006/taap.1993.1140
  16. Obremski K. Changes in Th1 and Th2 cytokine concentrations in ileal Peyer's patches in gilts exposed to zearalenone. Pol J Vet Sci 2014;17:53-9. https://doi.org/10.2478/pjvs-2014-0007
  17. Maresca M, Fantini J. Some food-associated mycotoxins as potential risk factors in humans predisposed to chronic intestinal inflammatory diseases. Toxicon 2010;56:282-94. https://doi.org/10.1016/j.toxicon.2010.04.016
  18. Przybylska-Gornowicz B, Lewczuk B, Prusik M, et al. The effects of deoxynivalenol and zearalenone on the pig large intestine. A light and electron microscopy study. Toxins 2018; 10:148. https://doi.org/10.3390/toxins10040148
  19. Przybylska-Gornowicz B, Tarasiuk M, Lewczuk B, et al. The effects of low doses of two Fusarium toxins, zearalenone and deoxynivalenol, on the pig jejunum. A light and electron microscopic study. Toxins 2015;7:4684-705. https://doi.org/10.3390/toxins7114684
  20. Cheng Q, Jiang S, Huang L, Ge J, Wang Y, Yang W. Zearalenone induced oxidative stress in the jejunum in postweaning gilts through modulation of the Keap1-Nrf2 signaling pathway and relevant genes. J Anim Sci 2019;97:1722-33. https://doi.org/10.1093/jas/skz051
  21. Xiao D, Yuan D, Tan B, Wang J, Liu Y, Tan B. The role of Nrf2 signaling pathway in Eucommia ulmoides flavones regulating oxidative stress in the intestine of piglets. Oxid Med Cell Longev 2019;2019:9719618. https://doi.org/10.1155/2019/9719618
  22. Kim SK, Yang JW, Kim MR, et al. Increased expression of Nrf2/ARE-dependent anti-oxidant proteins in tamoxifenresistant breast cancer cells. Free Radic Biol Med 2008;45: 537-46. https://doi.org/10.1016/j.freeradbiomed.2008.05.011
  23. Jiang SZ, Yang ZB, Yang WR, et al. Effects of purified zearalenone on growth performance, organ size, serum metabolites, and oxidative stress in postweaning gilts. J Anim Sci 2011;89: 3008-15. https://doi.org/10.2527/jas.2010-3658
  24. Jiang SZ, Yang ZB, Yang WR, et al. Effects of feeding purified zearalenone contaminated diets with or without clay enterosorbent on growth, nutrient availability, and genital organs in post-weaning female pigs. Asian-Australas J Anim Sci 2010;23:74-81. https://doi.org/10.5713/ajas.2010.90242
  25. Yang L, Wang S, Yang WR, et al. Effects of zearalenone on production performance, serum antioxidant capacity and immune function of weaning gilts. Chin J Anim Nutr 2017; 29:2843-50.
  26. Committee on Nutrient Requirements of Swine, National Research Council. Nutrient requirements of swine. 11th ed. Washington, DC, USA: National Academies Press; 2012.
  27. Latimer GW; AOAC International. Official methods of analysis of AOAC International. Gaithersburg, MD, USA: AOAC International; 2012.
  28. Jiang SZ, Yang ZB, Yang WR, et al. Effect on hepatonephric organs, serum metabolites and oxidative stress in post-weaning piglets fed purified zearalenone-contaminated diets with or without Calibrin-Z. J Anim Physiol Anim Nutr 2012;96:1147-56. https://doi.org/10.1111/j.1439-0396.2011.01233.x
  29. Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 1976;72:248-54. https://doi.org/10.1016/0003-2697(76)90527-3
  30. Zhou M, Yang L, Shao M, et al. Effects of zearalenone exposure on the TGF-β1/Smad3 signaling pathway and the expression of proliferation or apoptosis related genes of post-weaning gilts. Toxins 2018;10:49. https://doi.org/10.3390/toxins10020049
  31. Rivera A, Agnati LF, Horvath TL, Valderrama JJ, de La Calle A, Fuxe K. Uncoupling protein 2/3 immunoreactivity and the ascending dopaminergic and noradrenergic neuronal systems: relevance for volume transmission. Neuroscience 2006;137:1447-61. https://doi.org/10.1016/j.neuroscience.2005.05.051
  32. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method. Methods 2001;25:402-8. https://doi.org/10.1006/meth.2001.1262
  33. Tatay E, Espin S, Garcia-Fernandez AJ, Ruiz MJ. Oxidative damage and disturbance of antioxidant capacity by zearalenone and its metabolites in human cells. Toxicol In Vitro 2017;45: 334-9. https://doi.org/10.1016/j.tiv.2017.04.026
  34. Zheng W, Wang B, Si M, et al. Zearalenone altered the cytoskeletal structure via ER stress- autophagy- oxidative stress pathway in mouse TM4 Sertoli cells. Sci Rep 2018;8:3320. https://doi.org/10.1038/s41598-018-21567-8
  35. Bouaziz C, Sharaf el dein O, El Golli E, et al. Different apoptotic pathways induced by zearalenone, T-2 toxin and ochratoxin A in human hepatoma cells. Toxicology 2008;254:19-28. https://doi.org/10.1016/j.tox.2008.08.020
  36. Kouadio JH, Dano SD, Moukha S, Mobio TA, Creppy EE. Effects of combinations of Fusarium mycotoxins on the inhibition of macromolecular synthesis, malondialdehyde levels, DNA methylation and fragmentation, and viability in Caco-2 cells. Toxicon 2007;49:306-17. https://doi.org/10.1016/j.toxicon.2006.09.029
  37. Ren ZH, Deng HD, Wang YC, et al. The Fusarium toxin zearalenone and deoxynivalenol affect murine splenic antioxidant functions, interferon levels, and T-cell subsets. Environ Toxicol Pharmacol 2016;41:195-200. https://doi.org/10.1016/j.etap.2015.12.007
  38. Ren Z, Wang Y, Deng H, et al. Deoxynivalenol induces apoptosis in chicken splenic lymphocytes via the reactive oxygen species-mediated mitochondrial pathway. Environ Toxicol Pharmacol 2015;39:339-46. https://doi.org/10.1016/j.etap.2014.11.028
  39. Shi B, Su Y, Chang S, Sun Y, Meng X, Shan A. Vitamin C protects piglet liver against zearalenone-induced oxidative stress by modulating expression of nuclear receptors PXR and CAR and their target genes. Food Funct 2017;8:3675-87. https://doi.org/10.1039/C7FO01301A
  40. Cheng Q, Jiang S, Huang L, et al. Effects of zearalenone-induced oxidative stress and Keap1-Nrf2 signaling pathway-related gene expression in the ileum and mesenteric lymph nodes of post-weaning gilts. Toxicology 2020;429:152337. https://doi.org/10.1016/j.tox.2019.152337
  41. Palliyaguru DL, Chartoumpekis DV, Wakabayashi N, et al. Withaferin a induces nrf2-dependent protection against liver injury: role of keap1-independent mechanisms. Free Radic Biol Med 2016;101:116-28. https://doi.org/10.1016/j.freeradbiomed.2016.10.003
  42. Kubo E, Chhunchha B, Singh P, Sasaki H, Singh DP. Sulforaphane reactivates cellular antioxidant defense by inducing Nrf2/ARE/Prdx6 activity during aging and oxidative stress. Sci Rep 2017;7:14130. https://doi.org/10.1038/s41598-017-14520-8
  43. Mine Y, Young D, Yang C. Antioxidative stress effect of phosphoserine dimers is mediated via activation of the Nrf2 signaling pathway. Mol Nutr Food Res 2015;59:303-14. https://doi.org/10.1002/mnfr.201400381
  44. Ye Y, Li J, Yuan Z. Effect of antioxidant vitamin supplementation on cardiovascular outcomes: a meta-analysis of randomized controlled trials. PLoS One 2013;8:e56803. https://doi.org/10.1371/journal.pone.0056803
  45. Xiao Y, Zhang Z, Fu Y, Shan H, Cui S, Wu J. GSTA3 regulates TGF-β1-induced renal interstitial fibrosis in NRK-52E cells as a component of the PI3K-Keap1/Nrf2 pathway. J Int Med Res 2019;47:5787-801. https://doi.org/10.1177/0300060519876796
  46. Shanmugam G, Challa AK, Litovsky SH, et al. Enhanced Keap1-Nrf2 signaling protects the myocardium from isoproterenol-induced pathological remodeling in mice. Redox Biol 2019;27:101212. https://doi.org/10.1016/j.redox.2019.101212
  47. Huang Y, Zhou F, Shen C, Wang H, Xiao Y. LBP reduces theinflammatory injuryof kidney in septic rat and regulates the Keap1-Nrf2/ARE signaling pathway. Acta Cir Bras 2019; 34:e20190010000003. https://doi.org/10.1590/s0102-865020190010000003
  48. Wu CT, Deng JS, Huang WC, Shieh PC, Chung MI, Huang GJ. Salvianolic acid C against acetaminophen-induced acute liver injury by attenuating inflammation, oxidative stress, and apoptosis through inhibition of the Keap1/Nrf2/HO-1 signaling. Oxid Med Cell Longev 2019;2019:9056845. https://doi.org/10.1155/2019/9056845