Skip to main content
PMC home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1995 Jul;96(1):301–308. doi: 10.1172/JCI118035

Dual inhibition of nitric oxide and prostaglandin production contributes to the antiinflammatory properties of nitric oxide synthase inhibitors.

D Salvemini 1, P T Manning 1, B S Zweifel 1, K Seibert 1, J Connor 1, M G Currie 1, P Needleman 1, J L Masferrer 1
PMCID: PMC185201  PMID: 7542281

Abstract

We have recently put forward the hypothesis that the dual inhibition of proinflammatory nitric oxide (NO) and prostaglandins (PG) may contribute to the antiinflammatory properties of nitric oxide synthase (NOS) inhibitors. This hypothesis was tested in the present study. A rapid inflammatory response characterized by edema, high levels of nitrites (NO2-, a breakdown product of NO), PG, and cellular infiltration into a fluid exudate was induced by the administration of carrageenan into the subcutaneous rat air pouch. The time course of the induction of inducible nitric oxide synthase (iNOS) protein in the pouch tissue was found to coincide with the production of NO2-. Dexamethasone inhibited both iNOS protein expression and NO2- synthesis in the fluid exudate (IC50 = 0.16 mg/kg). Oral administration of N-iminoethyl-L-lysine (L-NIL) or NG-nitro-L-arginine methyl ester (NO2Arg) not only blocked nitrite accumulation in the pouch fluid in a dose-dependent fashion but also attenuated the elevated release of PG. Finally, carrageenan administration produced a time-dependent increase in cellular infiltration into the pouch exudate that was inhibited by dexamethasone and NOS inhibitors. At early times, i.e., 6 h, the cellular infiltrate is composed primarily of neutrophils (98%). Pretreatment with colchicine reduced both neutrophil infiltration and leukotriene B4 accumulation in the air pouch by 98% but did not affect either NO2- or PG levels. In conclusion, the major findings of this paper are that (a) selective inhibitors of iNOS are clearly antiinflammatory agents by inhibiting not only NO but also PG and cellular infiltration and (b) that neutrophils are not responsible for high levels of NO and PG produced.

Full text

PDF
308

Images in this article

302Image
on p.302
302Image
on p.302
303Image
on p.303
303Image
on p.303
304Image
on p.304

Selected References

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

  1. Connor J. R., Manning P. T., Settle S. L., Moore W. M., Jerome G. M., Webber R. K., Tjoeng F. S., Currie M. G. Suppression of adjuvant-induced arthritis by selective inhibition of inducible nitric oxide synthase. Eur J Pharmacol. 1995 Jan 24;273(1-2):15–24. doi: 10.1016/0014-2999(94)00672-t. [DOI] [PubMed] [Google Scholar]
  2. Corbett J. A., Kwon G., Turk J., McDaniel M. L. IL-1 beta induces the coexpression of both nitric oxide synthase and cyclooxygenase by islets of Langerhans: activation of cyclooxygenase by nitric oxide. Biochemistry. 1993 Dec 21;32(50):13767–13770. doi: 10.1021/bi00213a002. [DOI] [PubMed] [Google Scholar]
  3. DeWitt D. L. Prostaglandin endoperoxide synthase: regulation of enzyme expression. Biochim Biophys Acta. 1991 May 8;1083(2):121–134. doi: 10.1016/0005-2760(91)90032-d. [DOI] [PubMed] [Google Scholar]
  4. Flynn B. L. Rheumatoid arthritis and osteoarthritis: current and future therapies. Am Pharm. 1994 Nov;NS34(11):31-7, 41-2. doi: 10.1016/s0160-3450(15)30253-1. [DOI] [PubMed] [Google Scholar]
  5. Fu J. Y., Masferrer J. L., Seibert K., Raz A., Needleman P. The induction and suppression of prostaglandin H2 synthase (cyclooxygenase) in human monocytes. J Biol Chem. 1990 Oct 5;265(28):16737–16740. [PubMed] [Google Scholar]
  6. Futaki N., Arai I., Hamasaka Y., Takahashi S., Higuchi S., Otomo S. Selective inhibition of NS-398 on prostanoid production in inflamed tissue in rat carrageenan-air-pouch inflammation. J Pharm Pharmacol. 1993 Aug;45(8):753–755. doi: 10.1111/j.2042-7158.1993.tb07103.x. [DOI] [PubMed] [Google Scholar]
  7. Futaki N., Yoshikawa K., Hamasaka Y., Arai I., Higuchi S., Iizuka H., Otomo S. NS-398, a novel non-steroidal anti-inflammatory drug with potent analgesic and antipyretic effects, which causes minimal stomach lesions. Gen Pharmacol. 1993 Jan;24(1):105–110. doi: 10.1016/0306-3623(93)90018-s. [DOI] [PubMed] [Google Scholar]
  8. Inoue T., Fukuo K., Morimoto S., Koh E., Ogihara T. Nitric oxide mediates interleukin-1-induced prostaglandin E2 production by vascular smooth muscle cells. Biochem Biophys Res Commun. 1993 Jul 15;194(1):420–424. doi: 10.1006/bbrc.1993.1836. [DOI] [PubMed] [Google Scholar]
  9. Knowles R. G., Salter M., Brooks S. L., Moncada S. Anti-inflammatory glucocorticoids inhibit the induction by endotoxin of nitric oxide synthase in the lung, liver and aorta of the rat. Biochem Biophys Res Commun. 1990 Nov 15;172(3):1042–1048. doi: 10.1016/0006-291x(90)91551-3. [DOI] [PubMed] [Google Scholar]
  10. Masferrer J. L., Reddy S. T., Zweifel B. S., Seibert K., Needleman P., Gilbert R. S., Herschman H. R. In vivo glucocorticoids regulate cyclooxygenase-2 but not cyclooxygenase-1 in peritoneal macrophages. J Pharmacol Exp Ther. 1994 Sep;270(3):1340–1344. [PubMed] [Google Scholar]
  11. Masferrer J. L., Seibert K. Regulation of prostaglandin synthesis by glucocorticoids. Receptor. 1994 Spring;4(1):25–30. [PubMed] [Google Scholar]
  12. Masferrer J. L., Seibert K., Zweifel B., Needleman P. Endogenous glucocorticoids regulate an inducible cyclooxygenase enzyme. Proc Natl Acad Sci U S A. 1992 May 1;89(9):3917–3921. doi: 10.1073/pnas.89.9.3917. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Masferrer J. L., Zweifel B. S., Manning P. T., Hauser S. D., Leahy K. M., Smith W. G., Isakson P. C., Seibert K. Selective inhibition of inducible cyclooxygenase 2 in vivo is antiinflammatory and nonulcerogenic. Proc Natl Acad Sci U S A. 1994 Apr 12;91(8):3228–3232. doi: 10.1073/pnas.91.8.3228. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Masferrer J. L., Zweifel B. S., Seibert K., Needleman P. Selective regulation of cellular cyclooxygenase by dexamethasone and endotoxin in mice. J Clin Invest. 1990 Oct;86(4):1375–1379. doi: 10.1172/JCI114850. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Misko T. P., Schilling R. J., Salvemini D., Moore W. M., Currie M. G. A fluorometric assay for the measurement of nitrite in biological samples. Anal Biochem. 1993 Oct;214(1):11–16. doi: 10.1006/abio.1993.1449. [DOI] [PubMed] [Google Scholar]
  16. Moncada S., Higgs A. The L-arginine-nitric oxide pathway. N Engl J Med. 1993 Dec 30;329(27):2002–2012. doi: 10.1056/NEJM199312303292706. [DOI] [PubMed] [Google Scholar]
  17. Moncada S., Palmer R. M., Higgs E. A. Nitric oxide: physiology, pathophysiology, and pharmacology. Pharmacol Rev. 1991 Jun;43(2):109–142. [PubMed] [Google Scholar]
  18. Moore P. K., al-Swayeh O. A., Chong N. W., Evans R. A., Gibson A. L-NG-nitro arginine (L-NOARG), a novel, L-arginine-reversible inhibitor of endothelium-dependent vasodilatation in vitro. Br J Pharmacol. 1990 Feb;99(2):408–412. doi: 10.1111/j.1476-5381.1990.tb14717.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Moore W. M., Webber R. K., Jerome G. M., Tjoeng F. S., Misko T. P., Currie M. G. L-N6-(1-iminoethyl)lysine: a selective inhibitor of inducible nitric oxide synthase. J Med Chem. 1994 Nov 11;37(23):3886–3888. doi: 10.1021/jm00049a007. [DOI] [PubMed] [Google Scholar]
  20. Radomski M. W., Palmer R. M., Moncada S. Glucocorticoids inhibit the expression of an inducible, but not the constitutive, nitric oxide synthase in vascular endothelial cells. Proc Natl Acad Sci U S A. 1990 Dec;87(24):10043–10047. doi: 10.1073/pnas.87.24.10043. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Rees D. D., Cellek S., Palmer R. M., Moncada S. Dexamethasone prevents the induction by endotoxin of a nitric oxide synthase and the associated effects on vascular tone: an insight into endotoxin shock. Biochem Biophys Res Commun. 1990 Dec 14;173(2):541–547. doi: 10.1016/s0006-291x(05)80068-3. [DOI] [PubMed] [Google Scholar]
  22. Rettori V., Gimeno M., Lyson K., McCann S. M. Nitric oxide mediates norepinephrine-induced prostaglandin E2 release from the hypothalamus. Proc Natl Acad Sci U S A. 1992 Dec 1;89(23):11543–11546. doi: 10.1073/pnas.89.23.11543. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Salvemini D., Misko T. P., Masferrer J. L., Seibert K., Currie M. G., Needleman P. Nitric oxide activates cyclooxygenase enzymes. Proc Natl Acad Sci U S A. 1993 Aug 1;90(15):7240–7244. doi: 10.1073/pnas.90.15.7240. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Salvemini D., Seibert K., Masferrer J. L., Misko T. P., Currie M. G., Needleman P. Endogenous nitric oxide enhances prostaglandin production in a model of renal inflammation. J Clin Invest. 1994 May;93(5):1940–1947. doi: 10.1172/JCI117185. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Salvemini D., Settle S. L., Masferrer J. L., Seibert K., Currie M. G., Needleman P. Regulation of prostaglandin production by nitric oxide; an in vivo analysis. Br J Pharmacol. 1995 Mar;114(6):1171–1178. doi: 10.1111/j.1476-5381.1995.tb13330.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Sedgwick A. D., Lees P. Studies of eicosanoid production in the air pouch model of synovial inflammation. Agents Actions. 1986 Jun;18(3-4):429–438. doi: 10.1007/BF01965008. [DOI] [PubMed] [Google Scholar]
  27. Sedgwick A. D., Sin Y. M., Edwards J. C., Willoughby D. A. Increased inflammatory reactivity in newly formed lining tissue. J Pathol. 1983 Dec;141(4):483–495. doi: 10.1002/path.1711410406. [DOI] [PubMed] [Google Scholar]
  28. Seibert K., Masferrer J. L. Role of inducible cyclooxygenase (COX-2) in inflammation. Receptor. 1994 Spring;4(1):17–23. [PubMed] [Google Scholar]
  29. Vane J. R., Mitchell J. A., Appleton I., Tomlinson A., Bishop-Bailey D., Croxtall J., Willoughby D. A. Inducible isoforms of cyclooxygenase and nitric-oxide synthase in inflammation. Proc Natl Acad Sci U S A. 1994 Mar 15;91(6):2046–2050. doi: 10.1073/pnas.91.6.2046. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES