Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1996 Jan 9;93(1):255–259. doi: 10.1073/pnas.93.1.255

Molecular mechanisms of dexamethasone inhibition of nitric oxide synthase expression in interleukin 1 beta-stimulated mesangial cells: evidence for the involvement of transcriptional and posttranscriptional regulation.

D Kunz 1, G Walker 1, W Eberhardt 1, J Pfeilschifter 1
PMCID: PMC40217  PMID: 8552616

Abstract

Inducible nitric oxide synthase (iNOS; EC 1.14.13.39) is expressed in rat glomerular mesangial cells upon exposure to the inflammatory cytokine interleukin 1 beta (IL-1 beta). We have reported that nanomolar concentrations of dexamethasone suppress IL-1 beta-induced iNOS protein expression and production of nitrite, the stable end product of NO formation, without affecting IL-1 beta-triggered increase in iNOS mRNA levels. We now have studied the mechanisms by which dexamethasone suppresses IL-1 beta-stimulated iNOS expression in mesangial cells. Surprisingly, nuclear run-on transcription experiments demonstrate that dexamethasone markedly attenuates IL-1 beta-induced iNOS gene transcription. However, this is counteracted by a prolongation of the half-life of iNOS mRNA from 1 h to 2.5 h by dexamethasone. Moreover, dexamethasone drastically reduces the amount of iNOS protein by reduction of iNOS mRNA translation and increased degradation of iNOS protein. These results indicate that glucocorticoids act at multiple levels to regulate iNOS expression, thus providing important insights into the treatment of inflammatory diseases.

Full text

PDF
255

Images in this article

256Fig. 1
on p.256
257Fig. 3
on p.257
258Fig. 4
on p.258
258Fig. 5
on p.258

Selected References

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

  1. Anggård E. Nitric oxide: mediator, murderer, and medicine. Lancet. 1994 May 14;343(8907):1199–1206. doi: 10.1016/s0140-6736(94)92405-8. [DOI] [PubMed] [Google Scholar]
  2. Balligand J. L., Ungureanu-Longrois D., Simmons W. W., Pimental D., Malinski T. A., Kapturczak M., Taha Z., Lowenstein C. J., Davidoff A. J., Kelly R. A. Cytokine-inducible nitric oxide synthase (iNOS) expression in cardiac myocytes. Characterization and regulation of iNOS expression and detection of iNOS activity in single cardiac myocytes in vitro. J Biol Chem. 1994 Nov 4;269(44):27580–27588. [PubMed] [Google Scholar]
  3. Barnes P. J., Adcock I. Anti-inflammatory actions of steroids: molecular mechanisms. Trends Pharmacol Sci. 1993 Dec;14(12):436–441. doi: 10.1016/0165-6147(93)90184-l. [DOI] [PubMed] [Google Scholar]
  4. Beato M. Gene regulation by steroid hormones. Cell. 1989 Feb 10;56(3):335–344. doi: 10.1016/0092-8674(89)90237-7. [DOI] [PubMed] [Google Scholar]
  5. Beutler B., Krochin N., Milsark I. W., Luedke C., Cerami A. Control of cachectin (tumor necrosis factor) synthesis: mechanisms of endotoxin resistance. Science. 1986 May 23;232(4753):977–980. doi: 10.1126/science.3754653. [DOI] [PubMed] [Google Scholar]
  6. Cetkovic-Cvrlje M., Sandler S., Eizirik D. L. Nicotinamide and dexamethasone inhibit interleukin-1-induced nitric oxide production by RINm5F cells without decreasing messenger ribonucleic acid expression for nitric oxide synthase. Endocrinology. 1993 Oct;133(4):1739–1743. doi: 10.1210/endo.133.4.7691579. [DOI] [PubMed] [Google Scholar]
  7. Di Rosa M., Radomski M., Carnuccio R., Moncada S. Glucocorticoids inhibit the induction of nitric oxide synthase in macrophages. Biochem Biophys Res Commun. 1990 Nov 15;172(3):1246–1252. doi: 10.1016/0006-291x(90)91583-e. [DOI] [PubMed] [Google Scholar]
  8. Dignam J. D., Lebovitz R. M., Roeder R. G. Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. Nucleic Acids Res. 1983 Mar 11;11(5):1475–1489. doi: 10.1093/nar/11.5.1475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Eberhardt W., Kunz D., Pfeilschifter J. Pyrrolidine dithiocarbamate differentially affects interleukin 1 beta- and cAMP-induced nitric oxide synthase expression in rat renal mesangial cells. Biochem Biophys Res Commun. 1994 Apr 15;200(1):163–170. doi: 10.1006/bbrc.1994.1429. [DOI] [PubMed] [Google Scholar]
  10. Jonat C., Rahmsdorf H. J., Park K. K., Cato A. C., Gebel S., Ponta H., Herrlich P. Antitumor promotion and antiinflammation: down-modulation of AP-1 (Fos/Jun) activity by glucocorticoid hormone. Cell. 1990 Sep 21;62(6):1189–1204. doi: 10.1016/0092-8674(90)90395-u. [DOI] [PubMed] [Google Scholar]
  11. Kanno K., Hirata Y., Imai T., Iwashina M., Marumo F. Regulation of inducible nitric oxide synthase gene by interleukin-1 beta in rat vascular endothelial cells. Am J Physiol. 1994 Dec;267(6 Pt 2):H2318–H2324. doi: 10.1152/ajpheart.1994.267.6.H2318. [DOI] [PubMed] [Google Scholar]
  12. Kern J. A., Lamb R. J., Reed J. C., Daniele R. P., Nowell P. C. Dexamethasone inhibition of interleukin 1 beta production by human monocytes. Posttranscriptional mechanisms. J Clin Invest. 1988 Jan;81(1):237–244. doi: 10.1172/JCI113301. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Knowles R. G., Moncada S. Nitric oxide synthases in mammals. Biochem J. 1994 Mar 1;298(Pt 2):249–258. doi: 10.1042/bj2980249. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kunz D., Mühl H., Walker G., Pfeilschifter J. Two distinct signaling pathways trigger the expression of inducible nitric oxide synthase in rat renal mesangial cells. Proc Natl Acad Sci U S A. 1994 Jun 7;91(12):5387–5391. doi: 10.1073/pnas.91.12.5387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kunz D., Walker G., Pfeilschifter J. Dexamethasone differentially affects interleukin 1 beta- and cyclic AMP-induced nitric oxide synthase mRNA expression in renal mesangial cells. Biochem J. 1994 Dec 1;304(Pt 2):337–340. doi: 10.1042/bj3040337. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Mukaida N., Morita M., Ishikawa Y., Rice N., Okamoto S., Kasahara T., Matsushima K. Novel mechanism of glucocorticoid-mediated gene repression. Nuclear factor-kappa B is target for glucocorticoid-mediated interleukin 8 gene repression. J Biol Chem. 1994 May 6;269(18):13289–13295. [PubMed] [Google Scholar]
  17. Mühl H., Kunz D., Pfeilschifter J. Expression of nitric oxide synthase in rat glomerular mesangial cells mediated by cyclic AMP. Br J Pharmacol. 1994 May;112(1):1–8. doi: 10.1111/j.1476-5381.1994.tb13019.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Mühl H., Kunz D., Rob P., Pfeilschifter J. Cyclosporin derivatives inhibit interleukin 1 beta induction of nitric oxide synthase in renal mesangial cells. Eur J Pharmacol. 1993 Nov 2;249(1):95–100. doi: 10.1016/0014-2999(93)90666-6. [DOI] [PubMed] [Google Scholar]
  19. Nathan C., Xie Q. W. Regulation of biosynthesis of nitric oxide. J Biol Chem. 1994 May 13;269(19):13725–13728. [PubMed] [Google Scholar]
  20. Perrella M. A., Yoshizumi M., Fen Z., Tsai J. C., Hsieh C. M., Kourembanas S., Lee M. E. Transforming growth factor-beta 1, but not dexamethasone, down-regulates nitric-oxide synthase mRNA after its induction by interleukin-1 beta in rat smooth muscle cells. J Biol Chem. 1994 May 20;269(20):14595–14600. [PubMed] [Google Scholar]
  21. Pfeilschifter J. Anti-inflammatory steroids inhibit cytokine induction of nitric oxide synthase in rat renal mesangial cells. Eur J Pharmacol. 1991 Mar 19;195(1):179–180. doi: 10.1016/0014-2999(91)90399-b. [DOI] [PubMed] [Google Scholar]
  22. Pfeilschifter J. Cross-talk between transmembrane signalling systems: a prerequisite for the delicate regulation of glomerular haemodynamics by mesangial cells. Eur J Clin Invest. 1989 Aug;19(4):347–361. doi: 10.1111/j.1365-2362.1989.tb00241.x. [DOI] [PubMed] [Google Scholar]
  23. Pfeilschifter J., Kunz D., Mühl H. Nitric oxide: an inflammatory mediator of glomerular mesangial cells. Nephron. 1993;64(4):518–525. doi: 10.1159/000187394. [DOI] [PubMed] [Google Scholar]
  24. Pfeilschifter J., Rob P., Mülsch A., Fandrey J., Vosbeck K., Busse R. Interleukin 1 beta and tumour necrosis factor alpha induce a macrophage-type of nitric oxide synthase in rat renal mesangial cells. Eur J Biochem. 1992 Jan 15;203(1-2):251–255. doi: 10.1111/j.1432-1033.1992.tb19854.x. [DOI] [PubMed] [Google Scholar]
  25. Pfeilschifter J., Schwarzenbach H. Interleukin 1 and tumor necrosis factor stimulate cGMP formation in rat renal mesangial cells. FEBS Lett. 1990 Oct 29;273(1-2):185–187. doi: 10.1016/0014-5793(90)81080-8. [DOI] [PubMed] [Google Scholar]
  26. Pfeilschifter J., Vosbeck K. Transforming growth factor beta 2 inhibits interleukin 1 beta- and tumour necrosis factor alpha-induction of nitric oxide synthase in rat renal mesangial cells. Biochem Biophys Res Commun. 1991 Mar 15;175(2):372–379. doi: 10.1016/0006-291x(91)91574-v. [DOI] [PubMed] [Google Scholar]
  27. 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]
  28. Robbins R. A., Springall D. R., Warren J. B., Kwon O. J., Buttery L. D., Wilson A. J., Adcock I. M., Riveros-Moreno V., Moncada S., Polak J. Inducible nitric oxide synthase is increased in murine lung epithelial cells by cytokine stimulation. Biochem Biophys Res Commun. 1994 Feb 15;198(3):835–843. doi: 10.1006/bbrc.1994.1119. [DOI] [PubMed] [Google Scholar]
  29. Vodovotz Y., Bogdan C., Paik J., Xie Q. W., Nathan C. Mechanisms of suppression of macrophage nitric oxide release by transforming growth factor beta. J Exp Med. 1993 Aug 1;178(2):605–613. doi: 10.1084/jem.178.2.605. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. van de Stolpe A., Caldenhoven E., Stade B. G., Koenderman L., Raaijmakers J. A., Johnson J. P., van der Saag P. T. 12-O-tetradecanoylphorbol-13-acetate- and tumor necrosis factor alpha-mediated induction of intercellular adhesion molecule-1 is inhibited by dexamethasone. Functional analysis of the human intercellular adhesion molecular-1 promoter. J Biol Chem. 1994 Feb 25;269(8):6185–6192. [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES