Advertisement

Molecular Medicine

, Volume 11, Issue 1–12, pp 30–38 | Cite as

Ochratoxin A-Induced Renal Cortex Fibrosis and Epithelial-to-Mesenchymal Transition: Molecular Mechanisms of Ochratoxin A-Injury and Potential Effects of Red Wine

  • Nicoletta Gagliano
  • Carlo Torri
  • Elena Donetti
  • Fabio Grizzi
  • Francesco Costa
  • Alberto A E Bertelli
  • Massimiliano Migliori
  • Cristina Filippi
  • Marzia Bedoni
  • Vincenzo Panichi
  • Luca Giovannini
  • Magda Gioia
Articles

Abstract

We characterized the effect of chronic ochratoxin A (OTA) on rat kidney cortex, analyzing collagen content and collagen turnover and the major markers of epithelial-to-mesenchymal transition (EMT), such as α-smooth muscle actin (αSMA), cadherins, and MMP-9. Because OTA nephrotoxicity is mediated by free radicals, we also investigated whether antioxidants in red wine provided protection for the kidney and attenuated OTA-induced EMT. Collagen content, determined by computerized analysis of Sirius red-stained kidney sections, increased in OTA, OTA-wine, and OTA-EtOH treated rats. In kidney cortex homogenates, COL-I and COL-III mRNA levels tended to rise in OTA treated rats, but were similar to CT after OTA-wine and OTA-EtOH administration. TIMP-1 gene expression was up-regulated in OTA, OTA-wine, and OTA-EtOH treated rats. LH2b mRNA/COL-I mRNA was significantly up-regulated in OTA-wine and OTA-EtOH treated rats, compared with CT and OTA alone. TGF-βl signaling tended to dominate after OTA, OTA-wine, and OTA-EtOH. MMP-1 protein levels were not affected. OTA induced proMMP-9 and αSMA overexpression, decreases of E-cadherin and N-cadherin, and DSC-2 up-regulation. OTA-wine caused a further, unexpected decrease of E- and N-cadherins and further up-regulation of OTA-induced DSC-2, while strongly reducing the OTA-induced increases of αSMA and proMMP-9. Posttranslational collagen modifications, such as decreased collagen degradation through MMP inhibition and increased collagen cross-links, seem to be key mechanisms leading to OTA-induced kidney cortex fibrosis. This mechanism was not affected by red wine in these conditions. Red wine seems to have some protective role against OTA-induced EMT, although without completely blocking the process and determining a condition in which abundant cells display an intermediate translational phenotype, but there are no αSMA or epithelial markers.

Notes

Acknowledgements

We thank Silvia Celon and Osvaldo De Negri for their technical support and J.D. Baggott for editing the text. This work was supported by a grant of the Italian Government (Ministero delle Politiche Agricole e Forestali, MPAF) Targeted Project.

References

  1. 1.
    Marquardt RR, Frohlich A. (1992) A review of recent advances in understanding ochratoxicosis. J. Anim. Sci. 70:3968–88.CrossRefGoogle Scholar
  2. 2.
    Walker R. (2002) Risk assessment of ochratoxin: current views of the European Scientific Committee on Food, the JECFA and the Codex Committee on Food Additives and Contaminants. Adv. Exp. Med. Biol. 504:249–55.CrossRefGoogle Scholar
  3. 3.
    O’Brien E, Dietrich DR. (2005) Ochratoxin A: the continuing enigma. Crit. Rev. Toxicol. 35:33–60.CrossRefGoogle Scholar
  4. 4.
    Petkova-Bocharova T, Chernozemski IN, Castegnaro M. (1988) Ochratoxin A in human blood in relation to Balkan endemic nephropathy and urinary system tumours in Bulgaria. Food Addit. Contam. 5:299–301.CrossRefGoogle Scholar
  5. 5.
    Vrabcheva T et al. (2004) Analysis of ochratoxin A in foods consumed by inhabitants from an area with Balkan endemic nephropathy: a 1 month follow-up study. J. Agric. Food. Chem. 52:2404–10.CrossRefGoogle Scholar
  6. 6.
    Pfohl-Leszkowicz A, Petkova-Bocharova T, Chermozemsky IN, Castegnaro M. (2002) Balkan endemic nephropathy and associated urinary tract tumours: a review on aetiological causes and the potential role of mycotoxins. Food. Addit. Contam. 19:282–302.CrossRefGoogle Scholar
  7. 7.
    Krogh P. (1992) Role of ochratoxin in disease causation. Food. Chem. Toxicol. 30:213–24.CrossRefGoogle Scholar
  8. 8.
    Gekle M, Silbernagl S. (1996) Renal toxicodynamics of ochratoxin A: a pathophysiological approach. Kidney Blood Press. Res. 19:225–35.CrossRefGoogle Scholar
  9. 9.
    Klahr S, Schreiner G, Ichikawa I. (1988) The progression of renal disease. N. Engl. J. Med. 318:1657–66.CrossRefGoogle Scholar
  10. 10.
    Laurent GJ. (1987) Dynamic state of collagen degradation in vivo and their possible role in regulation of collagen mass. Am. J. Physiol. 252:C1–9.CrossRefGoogle Scholar
  11. 11.
    Lovett DH, Sterzel RB, Kashgarian M, Ryan JL. (1983) Neutral proteinases activity produced in vitro by cells of the glomerular mesangium. Kidney Int. 23:342–9.CrossRefGoogle Scholar
  12. 12.
    Sharma K, Ziyadeh FN. (1994) The emerging role of transforming growth factor-beta in kidney diseases (editorial). Am. J. Physiol. 266:F829–42.PubMedGoogle Scholar
  13. 13.
    Basile DP. (2001) Transforming growth factor-beta as a target for treatment in diabetic nephropathy. Am. J.Kidney Dis. 38:887–92.CrossRefGoogle Scholar
  14. 14.
    Liu Y. (2004) Hepatocyte growth factor in kidney fibrosis: therapeutic potential and mechanisms of action. Am. J. Physiol. Renal. Physiol. 287:F7–16.CrossRefGoogle Scholar
  15. 15.
    Mizuno S et al. (2000) Reciprocal balance of hepatocyte growth factor and transforming growth factor-beta 1 in renal fibrosis in mice. Kidney Int. 57: 937–48.CrossRefGoogle Scholar
  16. 16.
    Eddy AA. (1996) Molecular insights into renal interstitial fibrosis. J. Am. Soc. Nephrol. 7:2495–508.PubMedGoogle Scholar
  17. 17.
    Remuzzi G, Bertani T. (1998) Pathophysiology of progressive nephopathies. N. Engl. J. Med. 339:1448–56.CrossRefGoogle Scholar
  18. 18.
    Abrass CK et al. (1999) Unique changes in interstitial extracellular matrix composition are associated with rejection and cyclosporine toxicity in human renal allograft biopsies. Am. J. Kidney Dis. 33:11–20.CrossRefGoogle Scholar
  19. 19.
    Okada H, Danoff TM, Kalluri R, Neilson EG. (1997) Early role for Fsp1 in epithelialmesenchymal transformation. Am. J. Physiol. 273:F563–74.CrossRefGoogle Scholar
  20. 20.
    Powell DW et al. (1999) Myofibroblasts. I. Paracrine cells important in health and disease. Am. J. Physiol. 277:C1–9.CrossRefGoogle Scholar
  21. 21.
    Yang J, Lui J. (2002) Blockage of epithelial to myofibroblast transition by hepatocyte growth factor prevents renal interstitial fibrosis. J. Am. Soc. Nephrol. 13:96–107.PubMedGoogle Scholar
  22. 22.
    Yang J, Liu Y. (2001) Dissection of key events in tubular epithelial to myofibroblast transition and its implications in renal interstitial fibrosis. Am. J. Pathol. 159: 1465–75.CrossRefGoogle Scholar
  23. 23.
    Cheng S, Lovett DH. (2003) Gelatinase A (MMP-2) is necessary and sufficient for renal tubular cell epithelial-mesenchymal transformation. Am. J. Pathol. 162: 1937–49.CrossRefGoogle Scholar
  24. 24.
    Liu Y. (2005) Epithelial to mesenchymal transition in renal fibrogenesis: pathologic significance, molecular mechanisms, and therapeutic intervention. J. Am. Soc. Nephrol. 15:1–12.CrossRefGoogle Scholar
  25. 25.
    Baudrimont I et al. (1994) Effect of superoxide dismutase and catalase on the nephrotoxicity induced by subchronical administration of ochratoxin A in rats. Toxicology 89:101–11.CrossRefGoogle Scholar
  26. 26.
    Schaaf GJ et al. (2002) The role of oxidative stress in the ochratoxin A-mediated toxicity in proximal tubular cells. Biochim. Biophys. Acta 1588:149–58.CrossRefGoogle Scholar
  27. 27.
    Rodrigo R, Rivera GR. (2002) Renal damage mediated by oxidative stress: a hypothesis of protective effects of red wine. Free Rad. Biol. Med. 33:409–22.CrossRefGoogle Scholar
  28. 28.
    Rodrigo R et al. (2002) Rat kidney antioxidant response to long-term exposure to flavonol rich red wine. Life Sci. 71:2881–95.CrossRefGoogle Scholar
  29. 29.
    Bertelli AAE et al. (2005) Effect of ethanol and red wine on ochratoxin A-induced experimental acute nephrotoxicity. J. Agric. Food. Chem. 53:6924–9.CrossRefGoogle Scholar
  30. 30.
    Laemmli UK. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–5.CrossRefGoogle Scholar
  31. 31.
    Simon P, Godin M, Fillastre JP. (1996) Ochratoxin a: a new environmental factor which is toxic for the kidney? Nephrol. Dial. Transplant. 11:2389–91.CrossRefGoogle Scholar
  32. 32.
    Kane A, Creppy EE, Roschenthaler R, Dirheimer G. (1986) Changes in urinary and renal tubular enzymes caused by subchronic administration of ochratoxin A in rats. Toxicology 42:233–43.CrossRefGoogle Scholar
  33. 33.
    Gagliano N et al. (2000) Age-dependent expression of fibrosis-related genes and collagen deposition in rat kidney cortex. J. Gerontol. 55:B365–72.CrossRefGoogle Scholar
  34. 34.
    Steinmann-Niggli K, Ziswiler R, Kung M, Marti HP. (1998) Inhibition of metalloproteinases attenuates anti-Thy1.1 nephritis. J. Am. Soc. Nephrol. 9:397–407.PubMedGoogle Scholar
  35. 35.
    Sato Y, Fujimoto S, Hamai K, Eto T. (1998) Serial alterations of glomerular matrix-degrading metalloproteinase activity in anti-thymocyte-induced glomeru-lonephritis in rats. Nephron 78:195–200.CrossRefGoogle Scholar
  36. 36.
    Sakai T, Gross J. (1967) Some properties of the products of reaction of tadpole collagenase with collagen. Biochemistry 6:518–28.CrossRefGoogle Scholar
  37. 37.
    Walker LC, Overstreet MA, Yeowell HN. (2005) Tissue-specific expression and regulation of the alternatively-spliced forms of lysyl hydroxylase 2 (LH2) in human kidney cells and skin fibroblasts. Matrix Biol. 23:515–23.CrossRefGoogle Scholar
  38. 38.
    van der Slot AJ et al. (2004) Increased formation of pyridoline cross-links due to highetr telopeptide lysyl hydroxylase levels is a general fibrotic phenomenon. Matrix Biol. 23:251–7.CrossRefGoogle Scholar
  39. 39.
    Yu L, Border WA, Huang Y, Noble NA. (2003) TGF-β isoforms in renal fibrogenesis. Kidney Int. 64:844–56.CrossRefGoogle Scholar
  40. 40.
    German JB, Walzem RL. (2000) The health benefits of wine. Annu. Rev. Nutr. 20: 561–93.CrossRefGoogle Scholar
  41. 41.
    Abdel-Wahhab MA, Abdel-Galil MM, El-Lithey M. (2005) Melatonin counteracts oxidative stress in rats fed an ochratoxin A contaminated diet. J. Pineal Res. 38:130–5.CrossRefGoogle Scholar
  42. 42.
    Ozcelik N, Soyoz M, Kilink I. (2004) Effects of ochratoxin A on oxidative damage in rat kidney: protective role of melatonin. J. Appl. Toxicol. 24:211–5.CrossRefGoogle Scholar
  43. 43.
    Sauvant C, Holzinger H, Mildenberger S, Gekle M. (2005) Exposure to nephrotoxic ochratoxin A enhances collagen secretion in human renal proximal tubular cells. Mol. Nutr. Food. Res. 49:31–7.CrossRefGoogle Scholar
  44. 44.
    Zeisberg M, Kalluri R. (2004) The role of epithelial-to-mesenchymal transition in renal fibrosis. J. Mol. Med. 82:175–81.CrossRefGoogle Scholar
  45. 45.
    Masszi A et al. (2004) Integrity of cell-cell contacts is a critical regulator of TGF-β1-induced epithelial-to-myofibroblast transition. Am. J. Pathol. 165:1955–67.CrossRefGoogle Scholar
  46. 46.
    Nouwen EJ, Dauwe S, van der Biest I, De Broe ME. (1993) Stage- and segment-specific expression of cell-adhesion molecules N-CAM, A-CAM, and L-CAM in the kidney. Kidney Int. 44:147–58.CrossRefGoogle Scholar
  47. 47.
    Nadasdy T, Laszik Z, Blick KE, Johnson DL, Silva FG. (1994) Tubular atrophy in the end-stage kidney: a lectin and immunohistochemical study. Hum. Pathol. 25:22–8.CrossRefGoogle Scholar
  48. 48.
    Horster MF, Braun GS, Huber SM. (1999) Embryonic renal epithelia: induction, nephrogenesis, and cell differentiation. Physiol. Rev. 79:1157–91.CrossRefGoogle Scholar
  49. 49.
    Wheelock MJ, Johnson KR. (2003) Cadherins as modulators of cellular phenotype. Annu. Rev. Cell. Dev. Biol. 19:207–35.CrossRefGoogle Scholar
  50. 50.
    Collins JE et al. (1995) Regulation of desmocollin transcription in mouse preimplantation embryos. Development 121:743–53.PubMedGoogle Scholar
  51. 51.
    Matsumoto K, Nakamura T. (2001) Hepatocyte growth factor: renotropic role and potential therapeutics for renal diseases. Kidney Int. 59:2023–38.CrossRefGoogle Scholar

Copyright information

© Feinstein Institute for Medical Research 2005

Authors and Affiliations

  • Nicoletta Gagliano
    • 1
  • Carlo Torri
    • 1
  • Elena Donetti
    • 1
  • Fabio Grizzi
    • 2
  • Francesco Costa
    • 1
  • Alberto A E Bertelli
    • 1
  • Massimiliano Migliori
    • 3
  • Cristina Filippi
    • 3
  • Marzia Bedoni
    • 1
  • Vincenzo Panichi
    • 3
  • Luca Giovannini
    • 3
  • Magda Gioia
    • 1
  1. 1.Department of Human Morphology - LITA SegrateUniversity of MilanSegrate, MilanoItaly
  2. 2.Scientific DirectionIstituto Clinico Humanitas, Rozzano and “M. Rodriguez” Foundation, Institute for Quantitative Measures in MedicineMilanItaly
  3. 3.Department of Neuroscience (Pharmacology section) and Internal MedicineUniversity of PisaPisaItaly

Personalised recommendations