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Interleukin-6 changes tight junction permeability and intracellular phospholipid content in a human enterocyte cell culture model

Pediatric Surgery International volume 19, pages 321–325 (2003)Cite this article

Abstract.

Proinflammatory cytokines and secretory phospholipase A2 (sPLA2) are elevated in patients with inflammatory bowel disease (IBD). We previously reported that the proinflammatory cytokine IL-6 increased the expression of sPLA2 (a hydrolyzer of phosphatidylcholine) and decreased membrane integrity in an intestinal epithelial cell culture model. To determine the physiological effects of the IL-6 mediated increase in sPLA2 on decreased epithelial layer integrity, we investigated alterations of intracellular/secretory phospholipid (PL) composition in a cell culture model. In addition, since other PLs may also mediate epithelial membrane activity, we investigated the effect of IL-6 on PL activity in a Caco-2 enterocyte culture model. Caco-2 cells were incubated for 72 h with IL-6 or media alone (control). Both media and cell lysate were analyzed for PL composition using thin-layer chromatography. The PL composition in the media did not show any differences between the two groups (p>0.1). Total intracellular PL contents were also unchanged; however, IL-6 led to significant changes in PL composition including an increase in phosphatidylethanolamine (PE) and sphingomyelin (SM) and a decrease in phosphatidylcholine (PC) and lysophosphatidylcholine (LPC) (p<0.05). Both PE and SM are known as inflammatory signaling factors involved in human IBD. Our study suggests that the decreased membrane integrity seen with IL-6 application may occur via intracellular PL alterations, rather than through the direct effects of sPLA2.

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References

  1. Sartor RB (1994) Cytokines in intestinal inflammation: pathophysiological and clinical considerations. Gastroenterology 106:533–539

    Google Scholar 

  2. Sartor RB (1994) Cyclosporine therapy for inflammatory bowel disease. N Engl J Med 330:1897–1898

    Article  CAS  PubMed  Google Scholar 

  3. Reinecker HC, Steffen M, Witthoeft T, Pflueger I, Schreiber S, MacDermott RP, Raedler A (1993) Enhanced secretion of tumour necrosis factor-alpha, IL-6, and IL-1 beta by isolated lamina propria mononuclear cells from patients with ulcerative colitis and Crohn's disease. Clin Exp Immunol 94:174–181

    PubMed  Google Scholar 

  4. Gennari R, Alexander JW, Pyles T, Hartmann S, Ogle CK (1994) Effects of antimurine interleukin-6 on bacterial translocation during gut-derived sepsis. Arch Surg 129:1191–1197

    CAS  PubMed  Google Scholar 

  5. Kishikawa H, Miura S, Yoshida H, Hirokawa M, Nakamizo H, Higuchi H, Adachi M, Nakatsumi RC, Suzuki H, Saito H, Ishii H (2002) Transmural pressure induces IL-6 secretion by intestinal epithelial cells. Clin Exp Immunol 129:86–91

    Article  CAS  PubMed  Google Scholar 

  6. Molmenti EP, Ziambaras T, Perlmutter DH (1993) Evidence for an acute phase response in human intestinal epithelial cells. J Biol Chem 268:14116–14124

    CAS  PubMed  Google Scholar 

  7. Ziambaras T, Rubin DC, Perlmutter DH (1996) Regulation of sucrase-isomaltase gene expression in human intestinal epithelial cells by inflammatory cytokines. J Biol Chem 271:1237–1242

    Article  CAS  PubMed  Google Scholar 

  8. Vadas P, Scott K, Smith G, Rajkovic I, Stefanski E, Schouten BD, Singh R, Pruzanski W (1992) Serum phospholipase A2 enzyme activity and immunoreactivity in a prospective analysis of patients with septic shock. Life Sci 50:807–811

    Article  CAS  PubMed  Google Scholar 

  9. Vadas P (1984) Elevated plasma phospholipase A2 levels: correlation with the hemodynamic and pulmonary changes in gram-negative septic shock. J Lab Clin Med 104:873–881

    CAS  PubMed  Google Scholar 

  10. Otamiri T, Franzen L, Lindmark D, Tagesson C (1987) Increased phospholipase A2 and decreased lysophospholipase activity in the small intestinal mucosa after ischaemia and revascularisation. Gut 28:1445–1453

    CAS  PubMed  Google Scholar 

  11. Tagesson C, Telemo E, Ekstrom G, Westrom B (1987) Development of phospholipase A2 and lysophosphatidylcholine metabolising enzyme activities in the neonatal rat intestine. Gut 28:822–828

    CAS  PubMed  Google Scholar 

  12. Sawai T, Drongowski RA, Lampman RW, Coran AG, Harmon CM (2001) The effect of phospholipids and fatty acids on tight-junction permeability and bacterial translocation. Pediatr Surg Int 17:269–274

    Article  CAS  PubMed  Google Scholar 

  13. Holch JLM, Sloane Stanley GH (1959) A simple method for the isolation and purification of total lipids from animal tissues. J Biochem 226:497–509

    Google Scholar 

  14. Weinhold PAVC (1965) Phospholipid metabolism in the liver and lung of rats during development. Biochim Biophys Acta 106:540–550

    Article  CAS  PubMed  Google Scholar 

  15. Abe A, Gregory S, Lee L, Killen PD, Brady RO, Kulkarni A, Shayman JA (2000) Reduction of globotriaosylceramide in Fabry disease mice by substrate deprivation. J Clin Invest 105:1563–1571

    CAS  PubMed  Google Scholar 

  16. Brown KA, Back SJ, Ruchelli ED, Markowitz J, Mascarenhas M, Verma R, Piccoli DA, Baldassano RN (2002) Lamina propria and circulating interleukin-6 in newly diagnosed pediatric inflammatory bowel disease patients. Am J Gastroenterol 97:2603–2608

    Article  CAS  PubMed  Google Scholar 

  17. Murakami M, Kuwata H, Amakasu Y, Shimbara S, Nakatani Y, Atsumi G, Kudo I (1997) Prostaglandin E2 amplifies cytosolic phospholipase A2- and cyclooxygenase-2-dependent delayed prostaglandin E2 generation in mouse osteoblastic cells. Enhancement by secretory phospholipase A2. J Biol Chem 272:19891–19897

    Article  CAS  PubMed  Google Scholar 

  18. Balsinde J, Balboa MA, Li WH, Llopis J, Dennis EA (2000) Cellular regulation of cytosolic group IV phospholipase A2 by phosphatidylinositol bisphosphate levels. J Immunol 164:5398–5402

    CAS  PubMed  Google Scholar 

  19. Pruzanski W, Keystone EC, Sternby B, Bombardier C, Snow KM, Vadas P (1988) Serum phospholipase A2 correlates with disease activity in rheumatoid arthritis. J Rheumatol 15:1351–1355

    CAS  PubMed  Google Scholar 

  20. Minami T, Tojo H, Shinomura Y, Komatsubara T, Matsuzawa Y, Okamoto M (1993) Elevation of phospholipase A2 protein in sera of patients with Crohn's disease and ulcerative colitis. Am J Gastroenterol 88:1076–1080

    CAS  PubMed  Google Scholar 

  21. Minami T, Tojo H, Shinomura Y, Matsuzawa Y, Okamoto M (1994) Increased group II phospholipase A2 in colonic mucosa of patients with Crohn's disease and ulcerative colitis. Gut 35:1593–1598

    CAS  PubMed  Google Scholar 

  22. Minami T, Tojo H, Shinomura Y, Tarui S, Okamoto M (1992) Raised serum activity of phospholipase A2 immunochemically related to group II enzyme in inflammatory bowel disease: its correlation with disease activity of Crohn's disease and ulcerative colitis. Gut 33:914–921

    CAS  PubMed  Google Scholar 

  23. Yamashita S, Yamashita J, Ogawa M (1994) Overexpression of group II phospholipase A2 in human breast cancer tissues is closely associated with their malignant potency. Br J Cancer 69:1166–1170

    CAS  PubMed  Google Scholar 

  24. Minami T, Zushi S, Shinomura Y, Matsuzawa Y (1996) Phospholipase A2 stimulation of rat intestinal epithelial cell (IEC-6) migration. Am J Physiol 271:G664–G668

    CAS  PubMed  Google Scholar 

  25. Longo WE, Grossmann EM, Erickson B, Panesar N, Mazuski JE, Kaminski DL (1999) The effect of phospholipase A2 inhibitors on proliferation and apoptosis of murine intestinal cells. J Surg Res 84:51–56

    Article  CAS  PubMed  Google Scholar 

  26. Bertsch T, Banks RE, Forbes MA, Aufenanger J, Storr M, Illingworth JM, Perren TJ, Selby PJ, Kattermann R (1996) Phospholipase A2 activity in serum is induced during treatment with recombinant human interleukin-6 in patients with cancer. Ann Clin Biochem 33:565–567

    PubMed  Google Scholar 

  27. Crowl RM, Stoller TJ, Conroy RR, Stoner CR (1991) Induction of phospholipase A2 gene expression in human hepatoma cells by mediators of the acute phase response. J Biol Chem 266:2647–2651

    CAS  PubMed  Google Scholar 

  28. Homan R, Hamelehle KL (1998) Phospholipase A2 relieves phosphatidylcholine inhibition of micellar cholesterol absorption and transport by human intestinal cell line Caco-2. J Lipid Res 39:1197–1209

    CAS  PubMed  Google Scholar 

  29. Hannun YA (1994) The sphingomyelin cycle and the second messenger function of ceramide. J Biol Chem 269:3125–3128

    CAS  PubMed  Google Scholar 

  30. Kolesnick R (2002) The therapeutic potential of modulating the ceramide/sphingomyelin pathway. J Clin Invest 110:3–8

    Article  CAS  PubMed  Google Scholar 

  31. Dobrowsky RT, Kolesnick RN (2001) Analysis of sphingomyelin and ceramide levels and the enzymes regulating their metabolism in response to cell stress. Methods Cell Biol 66:135–165

    CAS  PubMed  Google Scholar 

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Affiliations

  1. Section of Pediatric Surgery, University of Michigan Medical School, F3970 Mott Children's Hospital, Ann Arbor, Michigan, 48109-0245, USA

    Y. Tazuke, R. A. Drongowski, D. H. Teitelbaum & A. G. Coran

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  1. Y. Tazuke
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  2. R. A. Drongowski
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  3. D. H. Teitelbaum
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  4. A. G. Coran
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Correspondence to A. G. Coran.

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Tazuke, Y., Drongowski, R.A., Teitelbaum, D.H. et al. Interleukin-6 changes tight junction permeability and intracellular phospholipid content in a human enterocyte cell culture model. Ped Surgery Int 19, 321–325 (2003). https://doi.org/10.1007/s00383-003-1003-8

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  • DOI: https://doi.org/10.1007/s00383-003-1003-8

Keywords

  • Proinflammatory cytokines
  • Interleukin-6
  • Secretory phospholipase A2
  • Phospholipid
  • Epithelial permeability
  • Inflammatory bowel disease