Skip to main content
SpringerLink
Log in

Phorbol myristate acetateex vivo model of enhanced colonic epithelial permeability

Reactive oxygen metabolite and protease independence

  • Intestinal Disorders, Inflammatory Bowel Disease, Irritable Bowel Syndrome, and Immunology
  • Published:
Digestive Diseases and Sciences Aims and scope Submit manuscript

Abstract

The initiating mechanisms involved in colonic injury are currently unknown. The goal of the current study was to examine the role of the inflammatory mediators reactive oxygen metabolites and proteases in anex vivo model of selective epithelial permeability. Rats were prepared with exteriorized colonic chambers to which the protein kinase C (PKC) activator phorbol myristate acetate (PMA) was added in doses ranging from 5 to 800 μg. PMA caused a dose-dependent transient increase in epithelial permeability, but had no significant effect on microvascular permeability. There was no accumulation of neutrophils and no apparent histological changes. PMA acts via a PKC-dependent mechanism, as assessed using the PKC-inactive phorbol analog 4α-phorbol didecanoate, and the response is tachyphylactic. The mechanism is independent of reactive oxygen metabolites and proteases, as shown by the lack of effect of the free radical scavengers superoxide dismutase and catalase and the general serine protease inhibitor soybean trypsin inhibitor. The classic inflammatory process does not appear to be involved in the PMA-induced epithelial permeability changes. This finding suggests that noninflammatory mechanisms may regulate the increased epithelial permeability induced by PMA. Further study to elucidate these mechanisms is of importance for understanding both normal gastrointestinal physiology and initiation of pathology.

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. Lewis KJ, Riddell RH, Weinstein WM: Gastrointestinal Pathology and Its Clinical Implications. Vol. II, New York, Igaku-Shoin, 1992

    Google Scholar 

  2. Hollander D, Vadheim CM, Brettholz E, Petersen GM, Delahunty T, Rotter JL: Increased intestinal permeability in patients with Crohn's disease and their relatives. A possible etiologic factor. Ann Intern Med 105:883–885, 1986

    Google Scholar 

  3. Hollander D: Permeability in Crohn's disease: altered barrier functions in healthy relatives? Gastroenterology 104:1848–1873, 1993

    Google Scholar 

  4. May GR, Sutherland LR, Meddings JB: Is small intestinal permeability really increased in relatives of patients with Crohn's disease? Gastroenterology 104:1627–1632, 1993

    Google Scholar 

  5. Onderdonk AB, Barlett JG: The role of bacteria in experimental ulcerative colitis. Am J Clin Nutr 32:258–265, 1979

    Google Scholar 

  6. Yamada T, Marshall S, Specian RD, Grisham MB: A comparative analysis of two models of colitis in rats. Gastroenterology 102:1524–1534, 1992

    Google Scholar 

  7. Grisham MB, Volkmer C, Tso P, Yamada T: Metabolism of trinitrobenzene sulfonic acid by the rat colon produces reactive oxygen species. Gastroenterology 101:540–547, 1991

    Google Scholar 

  8. Yamamoto S, Kiyoto I, Aizu E, Nakadote T, Hosoda Y, Kato R: Differential inhibition by staurosporine, a potent protein kinase C inhibitor, of 12-O-tetradecanoyl phorbol-13-acetate-caused skin tumor promotion, epidermal ornithine decarboxylase induction, hyperplasia, and inflammation. Carcinogenesis 10:1315–1322, 1989.

    Google Scholar 

  9. McCall CE, Taylor RG, Cousart SL, Woodruff RD, Lewis JC, O'Flaherty JT: Pulmonary injury induced by phorbol myristate acetate following intravenous administration in rabbits: Acute respiratory distress followed by pulmonary interstitial pneumonitis and pulmonary fibrosis. Am J Pathol 111:258–262, 1983

    Google Scholar 

  10. Taylor RG, McCall CE, Thrall RS, Woodbridge RD, O'Flaherty JT: Histopathological features of phorbol myristate acetate-induced lung injury. Lab Invest 52:61–70, 1985

    Google Scholar 

  11. Fretland DJ, Widomski D, Tsai BS, Zemaitis JM, Levin S, Djuric SW, Shone RL, Gaginella TS: The effect of leukotriene-B4 receptor antagonist SC-41730, on acetic acid-induced colonic inflammation. Agents Actions 27:225–227, 1989.

    Google Scholar 

  12. Buell MG, Berin MC: Histopathological characterization of a phorbol ester induced model of acute colitis in the rat. Gastroenterology 102:A599, 1992

    Google Scholar 

  13. Hecht G, Robinson B, Koutsouris A: Reversible disassembly of an intestinal epithelial monolayer by prolonged exposure to phorbol ester. Am J Physiol 266:G214-G221, 1994

    Google Scholar 

  14. Lehrer RI, Cohen L: Receptor-mediated regulation of superoxide production in human neutrophils stimulated by phorbol myristate acetate. J Clin Invest 68:1314–1320, 1981

    Google Scholar 

  15. Myers MA, McPhair LC, Snyderman R: Redistribution of protein kinase C activity in human monocytes: Correlation with activation of the respiratory burst. J Immunol 135:3411–3416, 1985

    Google Scholar 

  16. English D, Roloff JS, Lukens JN: Chemotactic factor enhancement of superoxide release from fluoride and phorbol myristate acetate stimulated neutrophils. Blood 58:129–134, 1981

    Google Scholar 

  17. Keisari Y, Geva I, Flescher E, Goldin H, Lavie G: Stimulation of human monocyte oxidative burst and related cytotoxicity by tumor-promoting and non-tumor-promoting diterpene esters, indole alkaloids and polyacetate-type agents. Int J Cancer 36:467–472, 1985

    Google Scholar 

  18. Hoult JRS, Nourshargh S: Phorbol myristate acetate enhances human polymorphonuclear neutrophil release of granular enzymes but inhibits chemokinesis. Br J Pharmacol 86:533–537, 1985

    Google Scholar 

  19. Rouis M, Nigon F, Lafuma C, Hornebeck W, Chapman MJ: Expression of elastase activity by human monocyte-macrophages is modulated by cellular cholesterol content, inflammatory mediators, and phorbol myristate acetate. Arteriosclerosis 10:246–255, 1990

    Google Scholar 

  20. Mentzer SJ, Herrmann SH, Crimmins MAV, Burakoff SJ, Faller DV: The role of CD18 in phorbol ester-induced human monocyte-mediated cytotoxicity. Cell Immun 115:66–77, 1988

    Google Scholar 

  21. Haverstick DM, Sakai H, Gray LS: Lymphocyte adhesion can be regulated by cytoskeleton-associated, PMA-induced capping of surface receptors. Am J Physiol 262:C916-C926, 1992

    Google Scholar 

  22. McIntyre TM, Reinhold SL, Prescott SM, Zimmerman GA: Protein kinase C activity appears to be required for the synthesis of platelet-activating factor and leukotriene B4 by human neutrophils. J Biol Chem 262:15370–15376, 1987

    Google Scholar 

  23. Hayashi M, Imai Y, Oh-ishi S: Phorbol ester stimulates PAF synthesis via the activation of protein kinase C in rat leukocytes. Lipids 26:1054–1059, 1991

    Google Scholar 

  24. Chou CF, Omary MB: Phorbol acetate enhances the phosphorylation of cytokeratins 8 and 18 in human colonic epithelial cells. FEBS Lett 282:200–204, 1991

    Google Scholar 

  25. Friedman E, Verderame M, Winawer S, Pollack R: Actin cytoskeletal organization loss in the benign-to-malignant tumor transition in cultured human colonic epithelial cells. Cancer Res 44:3040–3050, 1984

    Google Scholar 

  26. Phillips TE, Wilson J: Signal transduction pathways mediating mucin secretion from intestinal goblet cells. Dig Dis Sci 38:1046–1054, 1993

    Google Scholar 

  27. Shuntoh H, Taniyama K, Tanaka C: Involvement of protein kinase C in the Ca2+-dependent vesicular release of GABA from central and enteric neurons of the guinea pig. Brain Res 483:384–388, 1989

    Google Scholar 

  28. Tanaka C, Taniyama K, Kusunoki M: A phorbol ester and A23187 act synergistically to release acetylcholine from the guinea pig ileum. FEBS Lett 175:165–169, 1984

    Google Scholar 

  29. Sasaguri T, Watson SP: Phorbol esters inhibit smooth muscle contractions through activation of Na+−K+-ATPase. Br J Pharmacol 99:237–242, 1990

    Google Scholar 

  30. Vilaseca J, Salas A, Guarner F, Rodriguez R, Malagelada JR: Participation of thromboxane and other eicosanoid synthesis in the course of experimental inflammatory colitis. Gastroenterology 98:269–277, 1990

    Google Scholar 

  31. Wallace JL: Release of platelet-activating factor (PAF) and accelerated healing induced by a PAF antagonist in an animal model of chronic colitis. Can J Physiol Pharmacol 66:422–425, 1988

    Google Scholar 

  32. Keshavarzian A, Haydek J, Zabihi R, Doria M, D'Astice M, Sorenson JRJ: Agents capable of eliminating reactive oxygen species: Catalase, WR-2721, or Cu(II)2(3,5-DIPS)4 decrease experimental colitis. Dig Dis Sci 37:1866–1873, 1992

    Google Scholar 

  33. Thomas TK, Will PC, Srivastava A, Wilson CL, Harbison H, Little J, Chesonis RS, Pignatello M, Schmolze, Symington J, Kilian PL, Thompson RC: Evaluation of an interleukin-1 receptor antagonist in the rat acetic acid-induced colitis model. Agents Actions 34:187–190, 1991

    Google Scholar 

  34. Wallace JL, Keenan CM: Leukotriene B4 potentiates colonic ulceration in the rat. Dig Dis Sci 35:622–629, 1990

    Google Scholar 

  35. Wallace JL, MacNaughton WK, Morris GP, Beck PL: Inhibition of leukotriene synthesis markedly accelerates healing in a rat model of inflammatory bowel disease. Gastroenterology 96:29–36, 1989

    Google Scholar 

  36. Hoshino H, Sugiyama S, Ohara A, Goto H, Tsukamoto Y, Ozawa T: Mechanism and prevention of chronic colonic inflammation with trinitrobenzene sulfonic acid in rats. Clin Exp Pharm Physiol 19:717–722, 1992

    Google Scholar 

  37. Eliakim R, Karmeli F, Okon E, Rachmilewitz: Ketotifen effectively prevents mucosal damage in experimental colitis. Gut 33:1498–1503, 1992

    Google Scholar 

  38. Wallace JL, Higa A, McKnight GW, MacIntyre DE: Prevention and reversal of experimental colitis by a monoclonal antibody which inhibits leukocyte adherence. Inflammation 16:343–354, 1992

    Google Scholar 

  39. von Ritter C, Sekizuka E, Grisham MB, Granger DN: The chemotactic peptideN-formyl-methionyl-leucyl-phenylalanine increases mucosal permeability in the distal ileum of the rat. Gastroenterology 95:651–655, 1988

    Google Scholar 

  40. Dinda PK,Buell MG, Beck IT: Role of oxygen derived free radicals in the ethanol-induced small intestinal injury. Gastroenterology 102:A208, 1992

    Google Scholar 

  41. Fretland DJ, Widomski DL, Levin S, Gaginella TS: Colonic inflammation in the rabbit induced by phorbol-12-myristate-13-acetate. Inflammation 14:143–150, 1990

    Google Scholar 

  42. Luck MS, Bass P: Effect of epidermal growth factor on experimental colitis in the rat. J Pharmacol Exp Ther 264:984–990, 1993

    Google Scholar 

  43. Buell MG, Berin MC: Neutrophil independence of the initiation of colonic injury: Comparison of results from three models of experimental colitis in the rat. Dig Dis Sci 39:2575–2588, 1994

    Google Scholar 

  44. Henrikson CK, Argenaio RA, Liacos JA, Khosia J: Morphologic and functional effects of bile salt on porcine colon during injury and repair. Lab Invest 60:72–87, 1989

    Google Scholar 

  45. Joris I, DeGirolami U, Wortham K, Majno G: Vascular labeling with Monastral Blue B. Stain Technol 57:177–183, 1982

    Google Scholar 

  46. von Ritter C, Grisham MB, Hollwarth M, Inauen W, Granger DN: Neutrophil-derived oxidants mediate formyl-methionylleucyl-phenylalanine-induced increases in mucosal permeability in rats. Gastroenterology 97:778–780, 1989

    Google Scholar 

  47. Pihan G, Regillo C, Szabo S: Free Radicals and Lipid Peroxidation in Ethanol- or aspirin-induced gastric mucosal injury. Dig Dis Sci 32:1395–1401, 1987

    Google Scholar 

  48. Kvietys PR, Twohig B, Danzell J, Specian RD: Ethanol-induced injury to the rat gastric mucosa: Role of neutrophils and xanthine oxidase-derived radicals. Gastroenterology 98:909–920, 1990

    Google Scholar 

  49. Andrews FJ, Malcontenti C, O'Brien PE: Sequence of gastric mucosal injury following ischemia and reperfusion. Role of reactive oxygen metabolites. Dig Dis Sci 37:1256–1361, 1992

    Google Scholar 

  50. Vaananen PM, Meddings JB, Wallace JL: Role of oxygen derived free radicals in indomethacin induced gastric injury. Am J Physiol 261:G470-G475, 1991

    Google Scholar 

  51. Von Ritter CBR, Granger DN: Neutrophilic proteases: Mediators of formyl-methionyl-leucyl-phenylalanine-induced ileitis in rats. Gastroenterology 97:605–609, 1989

    Google Scholar 

  52. Weiss SJ: Tissue destruction by neutrophils. N Engl J Med 320:365–376, 1989

    Google Scholar 

  53. Madara JL: Epithelia: Biologic principles of organization.In Textbook of Gastroenterology, Vol. 1 T Yamada, DH Alpers, C. Owyang, DW Powell, FE Silverstein (eds.). Philadelphia, JB Lippincott, 1991, pp 102–118

    Google Scholar 

  54. Gabler WL, Bullock WW, Creamer HR: Phorbol myristate acetate induction of chemotactic migration of human polymorphonuclear neutrophils. Inflammation 17:521–530, 1993

    Google Scholar 

  55. O'Flaherty JT, Cousart S, Lineberger AS, Bond E, Bass A, DeChatelet LR, Leake ES, McCall CE: Phorbol myristate acetate: in vivo effects upon neutrophils, platelets, and lung. Am J Pathol 101:79–91, 1980

    Google Scholar 

  56. Nash S, Stafford J, Madara JL: The selective and superoxide independent disruption of intestinal epithelial tight junctions during leukocyte transmigration. Lab Invest 59:531–537, 1988

    Google Scholar 

  57. Davis RH, Mathias JR: Phorbol esters induce retrograde myoelectric activity in rabbit ileumin vivo. Am J Physiol 257:G578-G583, 1989

    Google Scholar 

  58. Soll AH, Toomey M, Culp D, Shanahan F, Beaven MA: Modulation of histamine release from canine fundic mucosal mast cells. Am J Physiol 254:G40-G48, 1988

    Google Scholar 

  59. Parks DA, Bulkley GB, Granger DN, Hamilton SR, McCord JM: Ischemic injury in the cat small intestine: Role of superoxide radicals. Gastroenterology 82:9–15, 1982

    Google Scholar 

  60. Kubes P, Arfors KE, Granger DN: Platelet-activating factor induced mucosal dysfunction: Role of oxidants and granulocytes. Am J Physiol 260:G965-G971, 1991

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Departments of Physiology and Medicine, Queen's University, Kingston, Ontario, Canada

    M. Cecilia Berin MSc &  Mikael G. Buell MD, PhD

  2. Division of Gastroenterology, Gastrointestinal Diseases Research Unit, Hotel Dieu Hospital, K7L 5G2, Kingston, Ontario, Canada

    M. Cecilia Berin MSc &  Mikael G. Buell MD, PhD

Authors
  1. M. Cecilia Berin MSc
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Mikael G. Buell MD, PhD
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Cecilia Berin, M., Buell, M.G. Phorbol myristate acetateex vivo model of enhanced colonic epithelial permeability. Digest Dis Sci 40, 2268–2279 (1995). https://doi.org/10.1007/BF02209017

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02209017

Key Words

Search

Navigation