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Intestinal apoptotic changes linked to metabolic status in fasted and refed rats

  • Gastrointestinal Function
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Abstract

Intestinal apoptosis and expression of apoptosis inducers – the cytokines TNFα, TGFβ1 – and the intestinal transcription factor Cdx2, were studied according to two different metabolic and hormonal phases which characterize long-term fasting: the long period of protein sparing during which energy expenditure is derived from lipid oxidation (phase II), and the later phase characterized by a rise in body protein utilization and plasma corticosterone (phase III). Apoptosis was further studied in 2, 6, and 24 h refed rats. Morphological apoptotic events were observed by environmental and conventional scanning electron microscopy and a TUNEL test was used to characterize the final stages of apoptotic death. The gene and protein expressions of TNFα, TGFβ1, and Cdx2 were measured. Apoptotic events and TNFα, TGFβ1, and Cdx2 gene and protein expressions did not vary significantly during phase II as compared to the normally fed animals. However, a phase III fasting induced a delay in intestinal epithelial apoptosis, along with a 92, 58, and 25% decrease in TNFα, TGFβ1, and Cdx2 mRNAs, respectively. The amounts of TNFα, TGFβ1, and Cdx2 proteins decreased by 70, 36, and 25%, respectively. Apoptosis was restored rapidly after a 2 h refeeding following the phase III, accompanied by a significant increase in TNFα, TGFβ1, and Cdx2 mRNA and the protein levels, compared to the phase III fasting values. The concomitant decreases in cytokines and Cdx2 and in apoptotic cells during phase III suggest the preservation of enterocytes during this critical fasting period in order to optimize nutrient absorption as soon as food is available and thus, to rapidly restore body mass.

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Abbreviations

Cdx2:

Caudal-related homeodomain protein

RT-PCR:

Reverse transcription–polymerase chain reaction

TGFβ1:

Transforming growth factor β1

TNFα:

Tumor necrosis factor α

References

  1. Barnard JA, Beauchamp RD, Coffey RJ, Moses HL (1989) Regulation of intestinal epithelial cell growth by transforming growth factor type beta. Proc Natl Acad Sci USA 86(5):1578–1582

    PubMed  Google Scholar 

  2. Belkhou R, Bechet D, Cherel Y, Galluser M, Ferrara M, le Maho Y (1994) Effect of fasting and thyroidectomy on cysteine proteinase activities in liver and muscle. Biochim Biophys Acta 1199(2):195–201

    PubMed  Google Scholar 

  3. Bonhomme C, Duluc I, Martin E, Chawengsaksophak K, Chenard MP, Kedinger M, Beck F, Freund JN, Domon-Dell C (2003) The Cdx2 homeobox gene has a tumour suppressor function in the distal colon in addition to a homeotic role during gut development. Gut 52(10):1465–1471

    Article  PubMed  Google Scholar 

  4. Caderni G, Perrelli MG, Cecchini F, Tessitore L (2002) Enhanced growth of colorectal aberrant crypt foci in fasted/refed rats involves changes in TGFbeta1 and p21CIP expressions. Carcinogenesis 23(2):323–327

    Article  PubMed  Google Scholar 

  5. Challet E, Le Maho Y, Robin JP, Malan A, Cherel Y (1995) Involvement of corticosterone in the fasting-induced rise in protein utilization and locomotor activity. Pharmacol Biochem Behav 50:405–412

    Article  PubMed  Google Scholar 

  6. Chen RH, Su YH, Chuang RL, Chang TY (1998) Suppression of transforming growth factor-beta-induced apoptosis through a phosphatidylinositol 3-kinase/Akt-dependent pathway. Oncogene 17(15):1959–1968

    Article  PubMed  Google Scholar 

  7. Cherel Y, Robin JP, Le Maho Y (1988) Physiology and biochemistry of long-term fasting in birds. Can J Zool 66:159–166

    Google Scholar 

  8. Chomczynski P, Sacchi N (1987) Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162(1):156–159

    Article  PubMed  Google Scholar 

  9. Ciacci C, Lind SE, Podolsky DK (1993) Transforming growth factor beta regulation of migration in wounded rat intestinal epithelial monolayers. Gastroenterology 105(1):93–101

    PubMed  Google Scholar 

  10. Diab-Assef M, Reimund JM, Ezenfis J, Duclos B, Kedinger M, Foltzer-Jourdainne C (2002) The phosphodiesterase inhibitor, pentoxifylline, alters rat intestinal epithelial cell proliferation via changes in the expression of transforming growth factors. Scand J Gastroenterol 37(2):206–214

    Article  PubMed  Google Scholar 

  11. Dignass AU, Podolsky DK (1993) Cytokine modulation of intestinal epithelial cell restitution: central role of transforming growth factor beta. Gastroenterology 105(5):1323–1332

    PubMed  Google Scholar 

  12. Domon-Dell C, Wang Q, Kim S, Kedinger M, Evers BM, Freund JN (2002) Stimulation of the intestinal Cdx2 homeobox gene by butyrate in colon cancer cells. Gut 50(4):525–529

    Article  PubMed  Google Scholar 

  13. Dunel-Erb S, Chevalier C, Laurent P, Bach A, Decrock F, Le Maho Y (2001) Restoration of the jejunal mucosa in rats refed after prolonged fasting. Comp Biochem Physiol 129A:933–947

    Google Scholar 

  14. Elitsur Y, Lichtman SN, Neace C, Dosescu J, Moshier JA (1998) Immunosuppressive effect of budesonide on human lamina propria lymphocytes. Immunopharmacology 38(3):279–285

    Article  PubMed  Google Scholar 

  15. Fesus L (1993) Biochemical events in naturally occurring forms of cell death. FEBS Lett 328:1–5

    Article  PubMed  Google Scholar 

  16. Fiocchi C (1997) Intestinal inflammation: a complex interplay of immune and nonimmune cell interactions. Am J Physiol 273(4 Pt 1):G769–G775

    PubMed  Google Scholar 

  17. Foligne B, Aissaoui S, Senegas-Balas F, Cayuela C, Bernard P, Antoine JM, Balas D (2001) Changes in cell proliferation and differentiation of adult rat small intestine epithelium after adrenalectomy: kinetic, biochemical, and morphological studies. Dig Dis Sci 46(6):1236–1246

    Article  PubMed  Google Scholar 

  18. Freund JN, Domon-Dell C, Kedinger M, Duluc I (1998) The Cdx-1 and Cdx-2 homeobox genes in the intestine. Biochem Cell Biol 76(6):957–969

    Article  PubMed  Google Scholar 

  19. Fukuyama K, Iwakiri R, Noda T, Kojima M, Utsumi H, Tsunada S, Sakata H, Ootani A, Fujimoto K (2001) Apoptosis induced by ischemia – reperfusion and fasting in gastric mucosa compared to small intestinal mucosa in rats. Dig Dis Sci 46(3):545–549

    Article  PubMed  Google Scholar 

  20. Geyra A, Uni Z, Gal-Garber O, Guy D, Sklan D (2002) Starving affects CDX gene expression during small intestinal development in the chick. J Nutr 132(5):911–917

    PubMed  Google Scholar 

  21. Goodman MN, Larsen PR, Kaplan MN, Aoki TT, Young VR, Ruderman NB (1980) Starvation in the rat. II. Effect of age and obesity on protein sparing and fuel metabolism. Am J Physiol 239:E277–E286

    PubMed  Google Scholar 

  22. Habold C, Chevalier C, Dunel-Erb S, Foltzer-Jourdainne C, Le Maho Y, Lignot JH (2004) Effects of fasting and refeeding on jejunal morphology and cellular activity in rats in relation to body stores depletion. Scand J Gastroenterol 39(6):531–539

    Article  PubMed  Google Scholar 

  23. Habold C, Foltzer-Jourdainne C, Le Maho Y, Lignot JH, Oudart H (2005) Intestinal gluconeogenesis and glucose transport according to body fuel availability in rats. J Physiol 566(Pt 2):575–586

    Article  PubMed  Google Scholar 

  24. Hague A, Bracey TS, Hicks DJ, Reed JC, Paraskeva C (1998) Decreased levels of p26-Bcl-2, but not p30 phosphorylated Bcl-2, precede TGFbeta1-induced apoptosis in colorectal adenoma cells. Carcinogenesis 19(9): 1691–1695

    Article  PubMed  Google Scholar 

  25. Hsu H, Shu HB, Pan MG, Goeddel DV (1996) TRADD-TRAF2 and TRADD-FADD interactions define two distinct TNF receptor 1 signal transduction pathways. Cell 26; 84(2):299–308

    Google Scholar 

  26. Ignotz RA, Massague J (1986) Transforming growth factor-beta stimulates the expression of fibronectin and collagen and their incorporation into the extracellular matrix. J Biol Chem 25; 261(9): 4337–4345

    Google Scholar 

  27. Iwakiri R, Gotoh Y, Noda T, Sugihara H, Fujimoto K, Fuseler J, Aw TY (2001) Programmed cell death in rat intestine: effect of feeding and fasting. Scand J Gastroenterol 36:39–47

    Article  PubMed  Google Scholar 

  28. Jeschke MG, Debroy MA, Wolf SE, Rajaraman S, Thompson JC (2000) Burn and starvation increase programmed cell death in small bowel epithelial cells. Dig Dis Sci 45(2):415–420

    Article  PubMed  Google Scholar 

  29. Kaiser GC, Polk DB (1997) Tumor necrosis factor alpha regulates proliferation in a mouse intestinal cell line. Gastroenterology 112(4):1231–1240

    Article  PubMed  Google Scholar 

  30. Koubi HE, Robin JP, Dewasmes G, Le Maho Y, Frutoso J, Minaire Y (1991) Fasting-induced rise in locomotor activity in rats coincides with increased protein utilization. Physiol Behav 50:337–343

    Article  PubMed  Google Scholar 

  31. Kurokowa M, Lynch K, Podolsky DK (1987) Effects of growth factors on an intestinal epithelial cell line: transforming growth factor beta inhibits proliferation and stimulates differentiation. Biochem Biophys Res Commun 13; 142(3):775–782

    Google Scholar 

  32. Le Maho Y, Vu Van Kha H, Koubi H, Dewasmes G, Girard J, Ferre P, Cagnard M (1981) Body composition, energy expenditure, and plasma metabolites in long-term fasting geese. Am J Physiol 41:E342–E354

    Google Scholar 

  33. Lorentz O, Duluc I, Arcangelis AD, Simon-Assmann P, Kedinger M, Freund JN (1997) Key role of the Cdx2 homeobox gene in extracellular matrix-mediated intestinal cell differentiation. J Cell Biol 15; 139(6):1553–1565

    Google Scholar 

  34. Luciano L, Gupta PD, Groos S, Adamski J (1995) Modulation of apoptosis by starvation: morphological and biochemical study of rat intestinal mucosa. Cell Death and Diff 2:259–266

    Google Scholar 

  35. Massague J (1996) TGFbeta signaling: receptors, transducers, and Mad proteins. Cell 28; 85(7):947–950

    Google Scholar 

  36. Oberhammer FA, Pavelka M, Sharma S, Tiefenbacher R, Purchio AF, Bursch W, Schulte-Hermann R (1992) Induction of apoptosis in cultured hepatocytes and in regressing liver by transforming growth factor beta 1. Proc Natl Acad Sci USA 15; 89(12):5408–5412

    Google Scholar 

  37. Piguet PF, Vesin C, Donati Y, Barazzone C (1999) TNF-induced enterocyte apoptosis and detachment in mice: induction of caspases and prevention by a caspase inhibitor, ZVAD-fmk. Lab Invest 79(4):495–500

    PubMed  Google Scholar 

  38. Roberts AB, Sporn MB (1993) Physiological actions and clinical applications of transforming growth factor-beta (TGF-beta). Growth Factors 8(1):1–9

    PubMed  Google Scholar 

  39. Ruemmele FM, Beaulieu JF, Dionne S, Levy E, Seidman EG, Cerf-Bensussan N, Lentze MJ (2002) Lipopolysaccharide modulation of normal enterocyte turnover by toll-like receptors is mediated by endogenously produced tumour necrosis factor alpha. Gut 51(6):842–848

    Article  PubMed  Google Scholar 

  40. Schaeffer C, Diab-Assef M, Plateroti M, Laurent-Huck F, Reimund JM, Kedinger M, Foltzer-Jourdainne C (2000) Cytokine gene expression during postnatal small intestinal development: regulation by glucocorticoids. Gut 47(2):192–198

    Article  PubMed  Google Scholar 

  41. Suh E, Chen L, Taylor J, Traber PG (1994) A homeodomain protein related to caudal regulates intestine-specific gene transcription. Mol Cell Biol 14(11):7340–7351

    PubMed  Google Scholar 

  42. Suh E, Traber PG (1996) An intestine-specific homeobox gene regulates proliferation and differentiation. Mol Cell Biol 16(2):619–625

    PubMed  Google Scholar 

  43. Trauth BC, Klas C, Peters AM, Matzku S, Moller P, Falk W, Debatin KM, Krammer PH (1989) Monoclonal antibody-mediated tumor regression by induction of apoptosis. Science 21; 245(4915):301–305

    Google Scholar 

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Acknowledgments

We thank M. Kedinger, C. Domon-Dell and J.N. Freund for their helpful discussion, and C. Arbiol and E. Martin for their technical help. We are also grateful for K.C. Flanders for providing us with the TGFβ1 antibody. C.H. was recipient of a Nestlé Nutrition grant. Support of the University Scientific Committee is gratefully acknowledged.

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Authors and Affiliations

  1. Centre d’Ecologie et Physiologie Energétiques, CNRS, 23 rue Becquerel, 67087, Strasbourg cedex 2, France

    Caroline Habold, Yvon Le Maho & Jean-Hervé Lignot

  2. INSERM, U381, 3 Avenue Molière, 67200, Strasbourg, France

    Charlotte Foltzer-Jourdainne

Authors
  1. Caroline Habold
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  2. Charlotte Foltzer-Jourdainne
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  3. Yvon Le Maho
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  4. Jean-Hervé Lignot
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Correspondence to Caroline Habold.

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Habold, C., Foltzer-Jourdainne, C., Le Maho, Y. et al. Intestinal apoptotic changes linked to metabolic status in fasted and refed rats. Pflugers Arch - Eur J Physiol 451, 749–759 (2006). https://doi.org/10.1007/s00424-005-1506-3

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  • DOI: https://doi.org/10.1007/s00424-005-1506-3

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