Deoxynivalenol inhibits the expression of trefoil factors (TFF) by intestinal human and porcine goblet cells

  • Fabien Graziani
  • Philippe Pinton
  • Hamza Olleik
  • Ange Pujol
  • Cendrine Nicoletti
  • Mehdi Sicre
  • Nathalie Quinson
  • El Hassan Ajandouz
  • Josette Perrier
  • Eric Di Pasquale
  • Isabelle P. OswaldEmail author
  • Marc MarescaEmail author
Organ Toxicity and Mechanisms


Trefoil factors (TFFs) are bioactive peptides expressed by several epithelia, including the intestine, where they regulate key functions such as tissue regeneration, barrier function and inflammation. Although food-associated mycotoxins, including deoxynivalenol (DON), are known to impact many intestinal functions, modulation of TFFs during mycotoxicosis has never been investigated. Here, we analyzed the effect of DON on TFFs expression using both human goblet cells (HT29-16E cells) and porcine intestinal explants. Results showed that very low doses of DON (nanomolar range) inhibit the secretion of TFFs by human goblet cells (IC50 of 361, 387 and 243 nM for TFF1, 2 and 3, respectively) and prevent wound healing. RT-qPCR analysis demonstrated that the inhibitory effect of DON is related to a suppression of TFFs mRNA expression. Experiments conducted on porcine intestinal explants confirmed the results obtained on cells. Finally, the use of specific inhibitors of signal pathways demonstrated that DON-mediated suppression of TFFs expression mainly involved Protein Kinase R and the MAP kinases (MAPK) p38 and ERK1/2. Taken together, our results show for the first time that at very low doses, DON suppresses the expression and production of intestinal TFFs and alters wound healing. Given the critical role of TFFs in tissue repair, our results suggest that DON-mediated suppression of TFFs contributes to the alterations of intestinal integrity the caused by this toxin.


Deoxynivalenol Mycotoxin Goblet cells TFF1 TFF2 TFF3 



We thank Dr Keda Tree for her help with the manuscript. We would like to thank Prof Laboisse and Dr Bou-Hanna who generously gave us the HT29-16E cells. We would also like to thank Dr Elise Courvoisier-Dezord in charge of the maintenance of the qPCR system at the AVB platform (iSm2, Marseille) and Anne Marie Cossalter (INRA, Toxalim, Toulouse) in charge of the animals. Finally, we thank Gaelle Le Flecher from Romer Labs France for her availability and valuable expertise on mycotoxins and their detection.

Author contributions

FG, AP, CN, HO, MS, NQ, EHA, ED, JP, MM performed experiments involving human cells. PP and IPO performed experiments involving porcine intestinal explants. IPO and MM supervised porcine and human experiments, respectively. IPO and MM wrote the paper.


This work was supported by the Centre National de la Recherche Scientifique (CNRS), the Institut National de la Recherche Agronomique (INRA), the Ministère de l’Enseignement Supérieur et de la Recherche Scientifique and the ANR Grants CaDON (ANR-15-CE21) and ExpoMycoPig (ANR-17-Carn012).

Compliance with ethical standards

Conflict of interest

The authors declare no conflict of interest.


  1. Aamann L, Vestergaard EM, Gronbaek H (2014) Trefoil factors in inflammatory bowel disease. World J Gastroenterol 20:3223–3230. CrossRefPubMedPubMedCentralGoogle Scholar
  2. Ajandouz EH, Berdah S, Moutardier V et al (2016) Hydrolytic fate of 3/15-acetyldeoxynivalenol in humans: specific deacetylation by the small intestine and liver revealed using in vitro and ex vivo approaches. Toxins (Basel) 8:232. CrossRefGoogle Scholar
  3. Alassane-Kpembi I, Puel O, Pinton P et al (2017) Co-exposure to low doses of the food contaminants deoxynivalenol and nivalenol has a synergistic inflammatory effect on intestinal explants. Arch Toxicol 91:2677–2687. CrossRefPubMedGoogle Scholar
  4. Beck PL, Wong JF, Li Y et al (2004) Chemotherapy- and radiotherapy-induced intestinal damage is regulated by intestinal trefoil factor. Gastroenterology 126:796–808. CrossRefPubMedGoogle Scholar
  5. Bennett JW, Klich M, Mycotoxins M (2003) Mycotoxins. Clin Microbiol Rev 16:497–516. CrossRefPubMedPubMedCentralGoogle Scholar
  6. Berthiller F, Crews C, Dall’asta C et al (2013) Masked mycotoxins: a review. Mol Nutr Food Res 57:165–186. CrossRefPubMedGoogle Scholar
  7. Chaiyarit P, Klanrit P, Photipakdee P et al (2015) Diagnostic value evaluation of trefoil factors family 3 for the early detection of colorectal cancer. World J Gastroenterol 18:1305–1312. Google Scholar
  8. Dellafiora L, Dall’Asta C, Galaverna G (2018) Toxicodynamics of mycotoxins in the framework of food risk assessment an in silico perspective. Toxins 10:52. CrossRefPubMedCentralGoogle Scholar
  9. Dignass A, Lynch-Devaney K, Kindon H et al (1994) Trefoil peptides promote epithelial migration through a transforming growth factor beta-independent pathway. J Clin Invest 94:376–383. CrossRefPubMedPubMedCentralGoogle Scholar
  10. Furuta GT, Turner JR, Taylor CT et al (2001) Hypoxia-inducible factor 1-dependent induction of intestinal trefoil factor protects barrier function during hypoxia. J Exp Med 193:1027–1034. CrossRefPubMedPubMedCentralGoogle Scholar
  11. García GRGR, Payros D, Pinton P et al (2018) Intestinal toxicity of deoxynivalenol is limited by Lactobacillus rhamnosus RC007 in pig jejunum explants. Arch Toxicol 92:983–993. CrossRefPubMedGoogle Scholar
  12. Gerding J, Cramer B, Humpf HU (2014) Determination of mycotoxin exposure in Germany using an LC-MS/MS multibiomarker approach. Mol Nutr Food Res 58:2358–2368. CrossRefPubMedGoogle Scholar
  13. Gourbeyre P, Berri M, Lippi Y et al (2015) Pattern recognition receptors in the gut: analysis of their expression along the intestinal tract and the crypt/villus axis. Physiol Rep 13:e12225. CrossRefGoogle Scholar
  14. Graziani F, Pujol A, Nicoletti C et al (2015) The food-associated ribotoxin deoxynivalenol modulates inducible NO synthase in human intestinal cell model. Toxicol Sci 145:372–382. CrossRefPubMedGoogle Scholar
  15. Hoffmann W (2005) Trefoil factors TFF (trefoil factor family) peptide-triggered signals promoting mucosal restitution. Cell Mol Life Sci 62:2932–2938. CrossRefPubMedGoogle Scholar
  16. Hoffmann W (2009) Trefoil factor family (TFF) peptides and chemokine receptors: a promising relationship. J Med Chem 52:6505–6510. CrossRefPubMedGoogle Scholar
  17. Kim YS, Ho SB (2010) Intestinal goblet cells and mucins in health and disease: recent insights and progress. Curr Gastroenterol Rep 12:319–330. CrossRefPubMedPubMedCentralGoogle Scholar
  18. Kjellev S (2009) The trefoil factor family—small peptides with multiple functionalities. Cell Mol Life Sci 66:1350–1369. CrossRefPubMedGoogle Scholar
  19. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-delta delta C(T)) method. Methods 25:402–408. CrossRefPubMedGoogle Scholar
  20. Lucioli J, Pinton P, Callu P et al (2013) The food contaminant deoxynivalenol activates the mitogen activated protein kinases in the intestine: Interest of ex vivo models as an alternative to in vivo experiments. Toxicon 66:31–36. CrossRefPubMedGoogle Scholar
  21. Madsen J, Nielsen O, Tornoe I et al (2007) Tissue localization of human trefoil factors 1, 2, and 3. J Histochem Cytochem 55:505–513. CrossRefPubMedGoogle Scholar
  22. Maresca M, Yahi N, Younes-Sakr L et al (2008) Both direct and indirect effects account for the pro-inflammatory activity of enteropathogenic mycotoxins on the human intestinal epithelium: stimulation of interleukin-8 secretion, potentiation of interleukin-1beta effect and increase in the transepithelial. Toxicol Appl Pharmacol 228:84–92. CrossRefPubMedGoogle Scholar
  23. Maresca M (2013) From the gut to the brain: journey and pathophysiological effects of the food-associated trichothecene mycotoxin deoxynivalenol. Toxins 5:784–820. CrossRefPubMedPubMedCentralGoogle Scholar
  24. Maresca M, Fantini J (2010) Some food-associated mycotoxins as potential risk factors in humans predisposed to chronic intestinal inflammatory diseases. Toxicon 56:282–294. CrossRefPubMedGoogle Scholar
  25. Mashimo H, Wu DC, Podolsky DK, Fishman MC (1996) Impaired defense of intestinal mucosa in mice lacking intestinal trefoil factor. Science 274:262–265. CrossRefPubMedGoogle Scholar
  26. Payros D, Alassane-Kpembi I, Pierron A et al (2016) Toxicology of deoxynivalenol and its acetylated and modified forms. Arch Toxicol 12:2931–2957. CrossRefGoogle Scholar
  27. Payros D, Dobrindt U, Martin P et al (2017) The food contaminant deoxynivalenol exacerbates the genotoxicity of gut microbiota. MBio 8:e00007–e17. CrossRefPubMedPubMedCentralGoogle Scholar
  28. Pestka JJ (2010) Deoxynivalenol: mechanisms of action, human exposure, and toxicological relevance. Arch Toxicol 84:663–679. CrossRefPubMedGoogle Scholar
  29. Pierron A, Mimoun S, Murate LS et al (2016a) Microbial biotransformation of DON: molecular basis for reduced toxicity. Sci Rep 6:29105. CrossRefPubMedPubMedCentralGoogle Scholar
  30. Pierron A, Mimoun S, Murate LS et al (2016b) Intestinal toxicity of the masked mycotoxin deoxynivalenol-3-beta-d-glucoside. Arch Toxicol 90:2037–2046. CrossRefPubMedGoogle Scholar
  31. Pierron A, Bracarense APFL, Cossalter AM et al (2018) Deepoxy-deoxynivalenol retains some immune-modulatory properties of the parent molecule deoxynivalenol in piglets. Arch Toxicol 92:3381–3389. CrossRefPubMedGoogle Scholar
  32. Pinton P, Oswald IP (2014) Effect of deoxynivalenol and other type B trichothecenes on the intestine: a review. Toxins (Basel) 6:1615–1643. CrossRefGoogle Scholar
  33. Pinton P, Braicu C, Nougayrede J-PP et al (2010) Deoxynivalenol impairs porcine intestinal barrier function and decreases the protein expression of claudin-4 through a mitogen-activated protein kinase-dependent mechanism. J Nutr 140:1956–1962. CrossRefPubMedGoogle Scholar
  34. Pinton P, Tsybulskyy D, Lucioli J et al (2012) Toxicity of deoxynivalenol and its acetylated derivatives on the intestine: differential effects on morphology, barrier function, tight junction proteins, and mitogen-activated protein kinases. Toxicol Sci 130:180–190. CrossRefPubMedGoogle Scholar
  35. Pinton P, Graziani F, Pujol A et al (2015) Deoxynivalenol inhibits the expression by goblet cells of intestinal mucins through a PKR and MAP kinase-dependent repression of the resistin-like molecule beta. Mol Nutr Food Res 59:1076–1087. CrossRefPubMedGoogle Scholar
  36. Playford RJ, Ghosh S, Mahmood A (2004) Growth factors and trefoil peptides in gastrointestinal health and disease. Curr Opin Pharmacol 4:567–571. CrossRefPubMedGoogle Scholar
  37. Potten CS, Kellett M, Roberts SA et al (1992) Measurement of in vivo proliferation in human colorectal mucosa using bromodeoxyuridine. Gut 33:71–78. CrossRefPubMedPubMedCentralGoogle Scholar
  38. Razafimanjato H, Garmy N, Guo X-J et al (2010) The food-associated fungal neurotoxin ochratoxin A inhibits the absorption of glutamate by astrocytes through a decrease in cell surface expression of the excitatory amino-acid transporters GLAST and GLT-1. Neurotoxicology 31:475–484. CrossRefPubMedGoogle Scholar
  39. Razafimanjato H, Benzaria A, Taieb N et al (2011) The ribotoxin deoxynivalenol affects the viability and functions of glial cells. Glia 59:1672–1683. CrossRefPubMedGoogle Scholar
  40. Robert H, Payros D, Pinton P et al (2017) Impact of mycotoxins on the intestine: are mucus and microbiota new targets? Crit Rev Toxicol 20:249–275. CrossRefGoogle Scholar
  41. Sturm A, Dignass AU (2008) Epithelial restitution and wound healing in inflammatory bowel disease. World J Gastroenterol 14:348–353. CrossRefPubMedPubMedCentralGoogle Scholar
  42. Taupin D, Podolsky DK (2003) Trefoil factors: initiators of mucosal healing. Nat Rev Mol Cell Biol 4:721–732. CrossRefPubMedGoogle Scholar
  43. Vandenbroucke V, Croubels S, Verbrugghe E et al (2009) The mycotoxin deoxynivalenol promotes uptake of Salmonella typhimurium in porcine macrophages, associated with ERK1/2 induced cytoskeleton reorganization. Vet Res 40:64. CrossRefPubMedGoogle Scholar
  44. Wan MLY, Turner PC, Allen KJ, El-Nezami H (2016) Lactobacillus rhamnosus GG modulates intestinal mucosal barrier and inflammation in mice following combined dietary exposure to deoxynivalenol and zearalenone. J Funct Foods 22:34–43. CrossRefGoogle Scholar
  45. Wu F, Groopman JD, Pestka JJ (2014) Public health impacts of foodborne mycotoxins. Annu Rev Food Sci Technol 5:351–372. CrossRefPubMedGoogle Scholar
  46. Xiao P, Ling H, Lan G et al (2015) Trefoil factors: gastrointestinal-specific proteins associated with gastric cancer. Clin Chim Acta 450:127–134. CrossRefPubMedGoogle Scholar
  47. Xie H, Guo J-H, An W-M et al (2017) Diagnostic value evaluation of trefoil factors family 3 for the early detection of colorectal cancer. World J Gastroenterol 23:2159–2167. CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.CNRS, Centrale Marseille, iSm2, UMR 7313Aix Marseille UniversityMarseilleFrance
  2. 2.Toxalim (Research Centre in Food Toxicology), INRA, ENVT, INP-Purpan, UPSUniversité de ToulouseToulouseFrance
  3. 3.CNRS, LNC UMR 7291Aix-Marseille UniversitéMarseilleFrance
  4. 4.CNRS, INP, Institute of NeurophysiopathologyAix-Marseille UniversityMarseilleFrance

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