Skip to main content
SpringerLink
Log in

Enterocyte response to ischemia is dependent on differentiation state

  • Published:
Journal of Gastrointestinal Surgery

Abstract

Enterocytes at the tips of microvilli are more sensitive to an ischemic insult than those cells residing in the crypts, an effect thought to be due to a relative lack of collateral flow. We speculated that this increased cellular sensitivity to ischemia might be an intrinsic feature of the cells related to their differentiated phenotype. To test this hypothesis, enterocyte response to ischemia was determined using both in vivo and in vitro models. For the in vivo studies, male Sprague-Dawley rats underwent laparotomy, and small intestinal ischemia was induced by clamping the superior mesenteric artery for 30 or 60 minutes, after which reperfusion was allowed for various time points up to 4 days. Injury was assessed histologically, as well as with Northern blots, probing for the enterocyte differentiation markers intestinal alkaline phosphatase and lactase, as well as the gut-epithelial marker villin. Mucosal changes consistent with ischemia/reperfusion injury were evident—that is, a rapid inflammatory response followed by progressive villus cell loss beginning at the tips and progressing to the crypts, depending on the degree of insult, with an eventual return to normal microanatomy. Intestinal alkaline phosphatase and lactase were lost immediately after ischemia and returned with reperfusion, confirming that the differentiated cells are particularly sensitive to ischemic injury. The in vitro studies employed two separate models of enterocyte differentiation: sodium butyrate-treated HT-29 cells and Caco-2 cells maintained for 7 days after confluence. In both models, undifferentiated and differentiated cells were subjected to treatment with 2-deoxyglucose and oligomycin-A (in vitro model of ischemia) and apoptosis was assessed by fluorescence-activated cell sorting analysis. Differentiation of both cell lines resulted in a significantly greater apoptotic response to ischemia compared to undifferentiated cells exposed to an identical insult. We conclude that differentiated enterocytes may be inherently more sensitive to ischemia-induced injury than their undifferentiated counterparts. These findings call into question the popularly held belief that villus tip cells are more susceptible to ischemia because of their location relative to the microvascular anatomy.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Potten CS. Epithelial cell growth and differentiation. II. Intestinal apoptosis. Am J Physiol 1997;273:253–257.

    Google Scholar 

  2. Gordon JI, Schmidt GH, Roth KA. Studies of intestinal stem cells using normal, chimeric and transgenic mice. FASEBJ 1992;6:3039–3050.

    CAS  Google Scholar 

  3. Traber PG. Diferentiation of intestinal epithelial cells: Lessons from the study of intestine-specific gene expression. J Lab ClinMed 1994;123:467–477.

    CAS  Google Scholar 

  4. Cheng H, Leblond CP. Origin, differentiation and renewal of the four main epithelial cell types in the mouse small intestine. V. Unitarian theory of the origin of the four epithelial cell types. AmJ Anat 1974;141:537–562.

    Article  CAS  Google Scholar 

  5. Gavriele Y, Sherman Y, Ben-Sasson SA. Identification of programmed cell death in-situ via specific labeling of nuclear DNA fragmentation. J Cell Biol 1992;119:493–501.

    Article  Google Scholar 

  6. Gordon JI. Intestinal epithelial differentiation: New insights from chimeric and transgenic mice. J Cell Biol 1989; 108:1187–1194.

    Article  PubMed  CAS  Google Scholar 

  7. Williamson RC. Intestinal adaptation (first of two parts). Structural, functional and cytokinetic changes. N Engl J Med 1978;298:1393–1402.

    Article  PubMed  CAS  Google Scholar 

  8. Williamson RC. Intestinal adaptation (second of two parts). Mechanisms of control. N Engl J Med 1978;298:1444–1450.

    Article  PubMed  CAS  Google Scholar 

  9. Mitsudo S, Brandt LJ. Pathology of intestinal ischemia. Surg Clin North Am 1992;72:43–63.

    PubMed  CAS  Google Scholar 

  10. Robinson JW, Mirkovitch V, Winistorfer B, Saegesser F. Response of the intestinal mucosa to ischemia. Gut 1981; 22:512–27.

    Article  PubMed  CAS  Google Scholar 

  11. Kampp M, Nilsson NJ. Extravascular shunting of oxygen in the small intestine of the cat. Acta Physiol Scand 1968;72:396–403.

    Article  PubMed  CAS  Google Scholar 

  12. Noda T, Iwarkiri R, Fujimoto K, Matsuo S, Aw JY. Programmed cell death induced by ischemia-reperfusion in rat intestinal small mucosa. Am J Physiol Gastrointest Liver Physiol 1998;274:270–276.

    Google Scholar 

  13. Ikeda H, Suzuki Y, Suzuki M, Koike M, Tamura J, Tong J, Nomura M, Itoh G. Apoptosis is a major mode of cell death caused by ischaemia and ischaemia/reperfusion injury to the rat intestinal epithelium. Gut 1998;42:530–537.

    Article  PubMed  CAS  Google Scholar 

  14. Sha KA, Green CJ. Characterization of apoptosis in intestinal ischaemia-reperfusion injury—a light and electron microscopic study. IntJ Exp Path 1997;78:355–363.

    Article  Google Scholar 

  15. Chirgwin JM, Przybla AE, MacDonald RJ, Rutter WJ. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry 1979;18:5294–5299.

    Article  PubMed  CAS  Google Scholar 

  16. Feinberg AP, Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Bioch 1983; 132:6–13.

    Article  CAS  Google Scholar 

  17. Henthorn PS, Raducha M, Edwards YH, Weiss MJ, Slaughter C, Lafferty MA, Harris H. Nucleotide and amino acid sequences of human intestinal alkaline phosphatase: Close homology to placental alkaline phosphatase. Proc Natl Acad Sci USA 1987;84:1234–1238.

    Article  PubMed  CAS  Google Scholar 

  18. Pringault E, Arpin M, Garcia A, Finidori J, Louvard D. A human villin cDNA clone to investigate the differentiation of intestinal and kidney cells in vivo and in culture. EMBO J 1986;5:3119–3124.

    PubMed  CAS  Google Scholar 

  19. Buller HA, Kothe MJC, Goldman DA, Grubman SA, Sasak WV, Matsudaira PT, Montgomery RK, Grand RJ. Coordinate expression of lactate-phlorizin hydrolase mRNA and enzyme levels in rat intestine during development. J Biol Chem 1990;265:6978–6983.

    PubMed  CAS  Google Scholar 

  20. Cleveland DW, Lopata MA, McDonald RJ, Conan NJ, Rutter WJ, Kirschner MW. Number and evolutionary conservation of α- and β-tubulin and cytoplasmic β- and γ-actin genes using specific cloned cDNA probes. Cell 1980;20:95–105.

    Article  PubMed  CAS  Google Scholar 

  21. Matthews JB, Smith JA, Tally KJ, Menconi MJ, Nguyen H, Fink MP. Chemical hypoxia increases junctional permeability and activates electrogenic ion transport in human epithelial monolayers. Surgery 1994;116:150–157.

    PubMed  CAS  Google Scholar 

  22. Hodin RA, Meng S, Archer S, Tang R. Cellular growth state differentially regulates enterocyte gene expression in butryate-treated HT-29 cells. Cell Growth Diff 1996;7:647–653.

    PubMed  CAS  Google Scholar 

  23. Pinto M, Robine-Leon S, Appay M, Kedinger M, Triadou N, Dussaulx E, Laeroix B, Simon-Assmann P, Maffen K, Fogh J, Zweibaum A. Enterocyte-like differentiation and polarization of the human colon carcinoma cell line Caco-2 in culture. Biol Cell 1983;47:323–330.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Harvard Medical School, Beth Israel Deaconess Medical Center/Massachusetts General Hospital, Boston, Massachusetts

    Brian F. Hinnebusch M.D., Qing Ma M.D., J. Welles Henderson B.A., Aleem Siddique M.D., Sonia Y. Archer M.D. & Richard A. Hodin M.D.

Authors
  1. Brian F. Hinnebusch M.D.
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Qing Ma M.D.
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. J. Welles Henderson B.A.
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Aleem Siddique M.D.
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Sonia Y. Archer M.D.
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Richard A. Hodin M.D.
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Richard A. Hodin M.D..

Additional information

Supported by grants DK47186, DK02527, and DK50623 from the National Institutes of Health.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Hinnebusch, B.F., Ma, Q., Henderson, J.W. et al. Enterocyte response to ischemia is dependent on differentiation state. J Gastrointest Surg 6, 403–409 (2002). https://doi.org/10.1016/S1091-255X(01)00076-2

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1016/S1091-255X(01)00076-2

Key words

Search

Navigation