Skip to main content
NCBI home page
Mediators of Inflammation logoLink to Mediators of Inflammation
. 1998;7(2):85–91. doi: 10.1080/09629359891225

The role of cyclooxygenase-1 and cyclooxygenase-2 in lipopolysaccharide and interleukin-1 stimulated enterocyte prostanoid formation.

W E Longo 1, L J Damore 1, J E Mazuski 1, G S Smith 1, N Panesar 1, D L Kaminski 1
PMCID: PMC1781826  PMID: 9836494

Abstract

Lipopolysaccharide is an inflammatory agent and interleukin-1 is a cytokine. Their pro-inflammatory effects may be mediated by prostanoids produced by inducible cyclooxygenase-2. The aim of this study was to determine the prostanoids produced by lipopolysaccharide and interleukin-1 stimulated enterocytes through the cyclooxygenase-1 and 2 pathways. Cultured enterocytes were stimulated with lipopolysaccharide or interleukin-1beta with and without cyclooxygenase inhibitors. Low concentrations of indomethacin and valerylsalicylic acid (VSA) were evaluated as cyclooxygenase-1 inhibitors and their effects compared with the effects of a specific cyclooxygenase-2 inhibitor, SC-58125. Prostaglandin E2, 6-keto prostaglandin F1alpha, prostaglandin D2 and leukotriene B4 levels were determined by radioimmunoassay. Immunoblot analysis using isoform-specific antibodies showed that the inducible cyclooxygenase enzyme (COX-2) was expressed by 4 h in LPS and IL-1beta treated cells while the constitutive COX-1 remained unaltered in its expression. Interleukin-1beta and lipopolysaccharide stimulated the formation of all prostanoids compared with untreated cells, but failed to stimulate leukotriene B4. Indomethacin at 20 microM concentration, and VSA inhibited lipopolysaccharide and interleukin 1beta stimulated prostaglandin E2, but not 6-keto prostaglandin F1alpha formation. SC-58125 inhibited lipopolysaccharide and interleukin-1beta stimulated 6-keto prostaglandin F1alpha but not prostaglandin E2 release. The specific cyclooxygenase-2 inhibitor also inhibited lipopolysaccharide produced prostaglandin D2 but not interleukin-1beta stimulated prostaglandin D2. While SC-58125 inhibited basal 6-keto prostaglandin-F1alpha formation it significantly increased basal prostaglandin E2 and prostaglandin D2 formation. As SC-58125 inhibited lipopolysaccharide and interleukin-1beta induced 6-keto prostaglandin F1alpha production but not prostaglandin E2 production, it suggests that these agents stimulate prostacyclin production through a cyclooxygenase-2 mediated mechanism and prostaglandin E2 production occurs through a cyclooxygenase-1 mediated mechanism. Prostaglandin D2 production appeared to be variably produced by cyclooxygenase-1 or cyclooxygenase-2, depending on the stimulus.

Full Text

The Full Text of this article is available as a PDF (213.0 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bhattacharyya D. K., Lecomte M., Dunn J., Morgans D. J., Smith W. L. Selective inhibition of prostaglandin endoperoxide synthase-1 (cyclooxygenase-1) by valerylsalicylic acid. Arch Biochem Biophys. 1995 Feb 20;317(1):19–24. doi: 10.1006/abbi.1995.1130. [DOI] [PubMed] [Google Scholar]
  2. Boolbol S. K., Dannenberg A. J., Chadburn A., Martucci C., Guo X. J., Ramonetti J. T., Abreu-Goris M., Newmark H. L., Lipkin M. L., DeCosse J. J. Cyclooxygenase-2 overexpression and tumor formation are blocked by sulindac in a murine model of familial adenomatous polyposis. Cancer Res. 1996 Jun 1;56(11):2556–2560. [PubMed] [Google Scholar]
  3. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  4. Capdevila J. H., Falck J. R., Estabrook R. W. Cytochrome P450 and the arachidonate cascade. FASEB J. 1992 Jan 6;6(2):731–736. doi: 10.1096/fasebj.6.2.1537463. [DOI] [PubMed] [Google Scholar]
  5. Cirino G. Multiple controls in inflammation. Extracellular and intracellular phospholipase A2, inducible and constitutive cyclooxygenase, and inducible nitric oxide synthase. Biochem Pharmacol. 1998 Jan 15;55(2):105–111. doi: 10.1016/s0006-2952(97)00215-3. [DOI] [PubMed] [Google Scholar]
  6. Deitch E. A., Beck S. C., Cruz N. C., De Maio A. Induction of heat shock gene expression in colonic epithelial cells after incubation with Escherichia coli or endotoxin. Crit Care Med. 1995 Aug;23(8):1371–1376. doi: 10.1097/00003246-199508000-00010. [DOI] [PubMed] [Google Scholar]
  7. DuBois R. N., Awad J., Morrow J., Roberts L. J., 2nd, Bishop P. R. Regulation of eicosanoid production and mitogenesis in rat intestinal epithelial cells by transforming growth factor-alpha and phorbol ester. J Clin Invest. 1994 Feb;93(2):493–498. doi: 10.1172/JCI116998. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Fong Y., Tracey K. J., Moldawer L. L., Hesse D. G., Manogue K. B., Kenney J. S., Lee A. T., Kuo G. C., Allison A. C., Lowry S. F. Antibodies to cachectin/tumor necrosis factor reduce interleukin 1 beta and interleukin 6 appearance during lethal bacteremia. J Exp Med. 1989 Nov 1;170(5):1627–1633. doi: 10.1084/jem.170.5.1627. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Futaki N., Takahashi S., Yokoyama M., Arai I., Higuchi S., Otomo S. NS-398, a new anti-inflammatory agent, selectively inhibits prostaglandin G/H synthase/cyclooxygenase (COX-2) activity in vitro. Prostaglandins. 1994 Jan;47(1):55–59. doi: 10.1016/0090-6980(94)90074-4. [DOI] [PubMed] [Google Scholar]
  10. Gierse J. K., Hauser S. D., Creely D. P., Koboldt C., Rangwala S. H., Isakson P. C., Seibert K. Expression and selective inhibition of the constitutive and inducible forms of human cyclo-oxygenase. Biochem J. 1995 Jan 15;305(Pt 2):479–484. doi: 10.1042/bj3050479. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hla T., Neilson K. Human cyclooxygenase-2 cDNA. Proc Natl Acad Sci U S A. 1992 Aug 15;89(16):7384–7388. doi: 10.1073/pnas.89.16.7384. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kargman S., Wong E., Greig G. M., Falgueyret J. P., Cromlish W., Ethier D., Yergey J. A., Riendeau D., Evans J. F., Kennedy B. Mechanism of selective inhibition of human prostaglandin G/H synthase-1 and -2 in intact cells. Biochem Pharmacol. 1996 Oct 11;52(7):1113–1125. doi: 10.1016/0006-2952(96)00462-5. [DOI] [PubMed] [Google Scholar]
  13. Lamura E., Hillier K., Kinkaid A., Wilton D. Compartmentalisation and characteristics of a Ca2+-dependent phospholipase A2 in human colon mucosa. Biochem Pharmacol. 1997 May 9;53(9):1323–1332. doi: 10.1016/s0006-2952(96)00883-0. [DOI] [PubMed] [Google Scholar]
  14. Lee P. P., Ip M. M. Regulation of proliferation of rat mammary tumor cells by inhibitors of cyclooxygenase and lipoxygenase. Prostaglandins Leukot Essent Fatty Acids. 1992 Jan;45(1):21–31. doi: 10.1016/0952-3278(92)90098-4. [DOI] [PubMed] [Google Scholar]
  15. Lyons-Giordano B., Pratta M. A., Galbraith W., Davis G. L., Arner E. C. Interleukin-1 differentially modulates chondrocyte expression of cyclooxygenase-2 and phospholipase A2. Exp Cell Res. 1993 May;206(1):58–62. doi: 10.1006/excr.1993.1120. [DOI] [PubMed] [Google Scholar]
  16. Meade E. A., Smith W. L., DeWitt D. L. Differential inhibition of prostaglandin endoperoxide synthase (cyclooxygenase) isozymes by aspirin and other non-steroidal anti-inflammatory drugs. J Biol Chem. 1993 Mar 25;268(9):6610–6614. [PubMed] [Google Scholar]
  17. Mitchell J. A., Akarasereenont P., Thiemermann C., Flower R. J., Vane J. R. Selectivity of nonsteroidal antiinflammatory drugs as inhibitors of constitutive and inducible cyclooxygenase. Proc Natl Acad Sci U S A. 1993 Dec 15;90(24):11693–11697. doi: 10.1073/pnas.90.24.11693. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Miyamoto T., Ogino N., Yamamoto S., Hayaishi O. Purification of prostaglandin endoperoxide synthetase from bovine vesicular gland microsomes. J Biol Chem. 1976 May 10;251(9):2629–2636. [PubMed] [Google Scholar]
  19. Morita I., Schindler M., Regier M. K., Otto J. C., Hori T., DeWitt D. L., Smith W. L. Different intracellular locations for prostaglandin endoperoxide H synthase-1 and -2. J Biol Chem. 1995 May 5;270(18):10902–10908. doi: 10.1074/jbc.270.18.10902. [DOI] [PubMed] [Google Scholar]
  20. Murakami M., Bingham C. O., 3rd, Matsumoto R., Austen K. F., Arm J. P. IgE-dependent activation of cytokine-primed mouse cultured mast cells induces a delayed phase of prostaglandin D2 generation via prostaglandin endoperoxide synthase-2. J Immunol. 1995 Nov 1;155(9):4445–4453. [PubMed] [Google Scholar]
  21. O'Sullivan M. G., Huggins E. M., Jr, Meade E. A., DeWitt D. L., McCall C. E. Lipopolysaccharide induces prostaglandin H synthase-2 in alveolar macrophages. Biochem Biophys Res Commun. 1992 Sep 16;187(2):1123–1127. doi: 10.1016/0006-291x(92)91313-f. [DOI] [PubMed] [Google Scholar]
  22. Peplow P. V. Actions of cytokines in relation to arachidonic acid metabolism and eicosanoid production. Prostaglandins Leukot Essent Fatty Acids. 1996 May;54(5):303–317. doi: 10.1016/s0952-3278(96)90044-7. [DOI] [PubMed] [Google Scholar]
  23. Rafferty J. F., Noguchi Y., Fischer J. E., Hasselgren P. O. Sepsis in rats stimulates cellular proliferation in the mucosa of the small intestine. Gastroenterology. 1994 Jul;107(1):121–127. doi: 10.1016/0016-5085(94)90069-8. [DOI] [PubMed] [Google Scholar]
  24. Salzman A. L., Wang H., Wollert P. S., Vandermeer T. J., Compton C. C., Denenberg A. G., Fink M. P. Endotoxin-induced ileal mucosal hyperpermeability in pigs: role of tissue acidosis. Am J Physiol. 1994 Apr;266(4 Pt 1):G633–G646. doi: 10.1152/ajpgi.1994.266.4.G633. [DOI] [PubMed] [Google Scholar]
  25. Sano H., Kawahito Y., Wilder R. L., Hashiramoto A., Mukai S., Asai K., Kimura S., Kato H., Kondo M., Hla T. Expression of cyclooxygenase-1 and -2 in human colorectal cancer. Cancer Res. 1995 Sep 1;55(17):3785–3789. [PubMed] [Google Scholar]
  26. Seibert K., Masferrer J. L. Role of inducible cyclooxygenase (COX-2) in inflammation. Receptor. 1994 Spring;4(1):17–23. [PubMed] [Google Scholar]
  27. Seibert K., Zhang Y., Leahy K., Hauser S., Masferrer J., Perkins W., Lee L., Isakson P. Pharmacological and biochemical demonstration of the role of cyclooxygenase 2 in inflammation and pain. Proc Natl Acad Sci U S A. 1994 Dec 6;91(25):12013–12017. doi: 10.1073/pnas.91.25.12013. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Sher M. E., D'Angelo A. J., Stein T. A., Bailey B., Burns G., Wise L. Cytokines in Crohn's colitis. Am J Surg. 1995 Jan;169(1):133–136. doi: 10.1016/s0002-9610(99)80121-4. [DOI] [PubMed] [Google Scholar]
  29. Simmons D. L., Levy D. B., Yannoni Y., Erikson R. L. Identification of a phorbol ester-repressible v-src-inducible gene. Proc Natl Acad Sci U S A. 1989 Feb;86(4):1178–1182. doi: 10.1073/pnas.86.4.1178. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Stratton M. D., Chandel B., Deshpande Y., Kaminski D. L., Li A. P., Vernava A. M., Longo W. E. The effect of Clostridium difficile toxin on colonocyte prostanoid activity. Prostaglandins. 1994 Dec;48(6):367–375. doi: 10.1016/0090-6980(94)90003-5. [DOI] [PubMed] [Google Scholar]
  31. Stratton M. D., Sexe R., Peterson B., Kaminski D. L., Li A. P., Longo W. E. The effect of trinitrobenzene sulfonic acid (TNB) on colonocyte arachidonic acid metabolism. J Surg Res. 1996 Feb 1;60(2):375–378. doi: 10.1006/jsre.1996.0061. [DOI] [PubMed] [Google Scholar]
  32. Williams C. S., DuBois R. N. Prostaglandin endoperoxide synthase: why two isoforms? Am J Physiol. 1996 Mar;270(3 Pt 1):G393–G400. doi: 10.1152/ajpgi.1996.270.3.G393. [DOI] [PubMed] [Google Scholar]
  33. Xie W. L., Chipman J. G., Robertson D. L., Erikson R. L., Simmons D. L. Expression of a mitogen-responsive gene encoding prostaglandin synthase is regulated by mRNA splicing. Proc Natl Acad Sci U S A. 1991 Apr 1;88(7):2692–2696. doi: 10.1073/pnas.88.7.2692. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Mediators of Inflammation are provided here courtesy of Wiley

RESOURCES