Skip to main content
PMC home page
British Journal of Pharmacology logoLink to British Journal of Pharmacology
. 1994 Jun;112(2):396–402. doi: 10.1111/j.1476-5381.1994.tb13085.x

Induction of nitric oxide synthase in cultured vascular smooth muscle cells: the role of cyclic AMP.

K Hirokawa 1, K O'Shaughnessy 1, K Moore 1, P Ramrakha 1, M R Wilkins 1
PMCID: PMC1910341  PMID: 7521256

Abstract

1. Interleukin-1 beta (IL-1 beta) is a potent stimulant of inducible nitric oxide synthase (iNOS) mRNA and nitric oxide (NO) production in vascular smooth muscle (VSM) cells in culture. These studies investigate the role of adenosine 3':5'-cyclic monophosphate (cyclic AMP) in this process. 2. Dibutyryl cyclic AMP (db cyclic AMP, 0.1-1 mM), forskolin (1-10 microM) and the phosphodiesterase inhibitor, Ro 20-1724 (1-10 microM), all of which increase intracellular cyclic AMP, had no effect on NO production when added alone but markedly enhanced NO production by IL-1 beta-stimulated VSM cells in a dose-dependent manner. Consistent with a cyclic AMP-mediated action, isoprenaline (1-10 microM) increased NO production from IL-1 beta-stimulated cells. Dibutyryl cyclic GMP (db cyclic GMP) had no effect at concentrations up to 1 mM. 3. Pursuing these observations, iNOS protein levels were examined by Western blot analysis and iNOS mRNA levels were measured by reverse transcription and amplification of the resultant cDNA using the polymerase chain reaction. In addition to enhancing NO production, db cyclic AMP increased iNOS protein and mRNA above that produced by IL-1 beta alone. 4. These data demonstrate a major effect of cyclic AMP on cytokine-induced NOS activity in VSM cells, mediated at least in part by regulating synthesis of iNOS, and has implications for the pathogenesis and management of septic shock.

Full text

PDF
402

Images in this article

400Figure 6
on p.400
400Figure 7
on p.400

Selected References

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

  1. Beasley D., Schwartz J. H., Brenner B. M. Interleukin 1 induces prolonged L-arginine-dependent cyclic guanosine monophosphate and nitrite production in rat vascular smooth muscle cells. J Clin Invest. 1991 Feb;87(2):602–608. doi: 10.1172/JCI115036. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bredt D. S., Ferris C. D., Snyder S. H. Nitric oxide synthase regulatory sites. Phosphorylation by cyclic AMP-dependent protein kinase, protein kinase C, and calcium/calmodulin protein kinase; identification of flavin and calmodulin binding sites. J Biol Chem. 1992 Jun 5;267(16):10976–10981. [PubMed] [Google Scholar]
  3. Brown L. A., Nunez D. J., Wilkins M. R. Differential regulation of natriuretic peptide receptor messenger RNAs during the development of cardiac hypertrophy in the rat. J Clin Invest. 1993 Dec;92(6):2702–2712. doi: 10.1172/JCI116887. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brüne B., Lapetina E. G. Phosphorylation of nitric oxide synthase by protein kinase A. Biochem Biophys Res Commun. 1991 Dec 16;181(2):921–926. doi: 10.1016/0006-291x(91)91279-l. [DOI] [PubMed] [Google Scholar]
  5. Busse R., Mülsch A. Induction of nitric oxide synthase by cytokines in vascular smooth muscle cells. FEBS Lett. 1990 Nov 26;275(1-2):87–90. doi: 10.1016/0014-5793(90)81445-t. [DOI] [PubMed] [Google Scholar]
  6. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  7. Dinerman J. L., Lowenstein C. J., Snyder S. H. Molecular mechanisms of nitric oxide regulation. Potential relevance to cardiovascular disease. Circ Res. 1993 Aug;73(2):217–222. doi: 10.1161/01.res.73.2.217. [DOI] [PubMed] [Google Scholar]
  8. Durieu-Trautmann O., Fédérici C., Créminon C., Foignant-Chaverot N., Roux F., Claire M., Strosberg A. D., Couraud P. O. Nitric oxide and endothelin secretion by brain microvessel endothelial cells: regulation by cyclic nucleotides. J Cell Physiol. 1993 Apr;155(1):104–111. doi: 10.1002/jcp.1041550114. [DOI] [PubMed] [Google Scholar]
  9. Feinstein D. L., Galea E., Reis D. J. Norepinephrine suppresses inducible nitric oxide synthase activity in rat astroglial cultures. J Neurochem. 1993 May;60(5):1945–1948. doi: 10.1111/j.1471-4159.1993.tb13425.x. [DOI] [PubMed] [Google Scholar]
  10. Fort P., Marty L., Piechaczyk M., el Sabrouty S., Dani C., Jeanteur P., Blanchard J. M. Various rat adult tissues express only one major mRNA species from the glyceraldehyde-3-phosphate-dehydrogenase multigenic family. Nucleic Acids Res. 1985 Mar 11;13(5):1431–1442. doi: 10.1093/nar/13.5.1431. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gaillard T., Mülsch A., Busse R., Klein H., Decker K. Regulation of nitric oxide production by stimulated rat Kupffer cells. Pathobiology. 1991;59(4):280–283. doi: 10.1159/000163663. [DOI] [PubMed] [Google Scholar]
  12. Gaillard T., Mülsch A., Klein H., Decker K. Regulation by prostaglandin E2 of cytokine-elicited nitric oxide synthesis in rat liver macrophages. Biol Chem Hoppe Seyler. 1992 Sep;373(9):897–902. doi: 10.1515/bchm3.1992.373.2.897. [DOI] [PubMed] [Google Scholar]
  13. Hortelano S., Genaro A. M., Boscá L. Phorbol esters induce nitric oxide synthase activity in rat hepatocytes. Antagonism with the induction elicited by lipopolysaccharide. J Biol Chem. 1992 Dec 15;267(35):24937–24940. [PubMed] [Google Scholar]
  14. Hortelano S., Genaro A. M., Boscá L. Phorbol esters induce nitric oxide synthase and increase arginine influx in cultured peritoneal macrophages. FEBS Lett. 1993 Apr 5;320(2):135–139. doi: 10.1016/0014-5793(93)80078-9. [DOI] [PubMed] [Google Scholar]
  15. Karpinski B. A., Morle G. D., Huggenvik J., Uhler M. D., Leiden J. M. Molecular cloning of human CREB-2: an ATF/CREB transcription factor that can negatively regulate transcription from the cAMP response element. Proc Natl Acad Sci U S A. 1992 Jun 1;89(11):4820–4824. doi: 10.1073/pnas.89.11.4820. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Koide M., Kawahara Y., Nakayama I., Tsuda T., Yokoyama M. Cyclic AMP-elevating agents induce an inducible type of nitric oxide synthase in cultured vascular smooth muscle cells. Synergism with the induction elicited by inflammatory cytokines. J Biol Chem. 1993 Nov 25;268(33):24959–24966. [PubMed] [Google Scholar]
  17. Marotta P., Sautebin L., Di Rosa M. Modulation of the induction of nitric oxide synthase by eicosanoids in the murine macrophage cell line J774. Br J Pharmacol. 1992 Nov;107(3):640–641. doi: 10.1111/j.1476-5381.1992.tb14499.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Michel T., Li G. K., Busconi L. Phosphorylation and subcellular translocation of endothelial nitric oxide synthase. Proc Natl Acad Sci U S A. 1993 Jul 1;90(13):6252–6256. doi: 10.1073/pnas.90.13.6252. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. 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]
  20. Nakane M., Mitchell J., Förstermann U., Murad F. Phosphorylation by calcium calmodulin-dependent protein kinase II and protein kinase C modulates the activity of nitric oxide synthase. Biochem Biophys Res Commun. 1991 Nov 14;180(3):1396–1402. doi: 10.1016/s0006-291x(05)81351-8. [DOI] [PubMed] [Google Scholar]
  21. Nakayama D. K., Geller D. A., Lowenstein C. J., Chern H. D., Davies P., Pitt B. R., Simmons R. L., Billiar T. R. Cytokines and lipopolysaccharide induce nitric oxide synthase in cultured rat pulmonary artery smooth muscle. Am J Respir Cell Mol Biol. 1992 Nov;7(5):471–476. doi: 10.1165/ajrcmb/7.5.471. [DOI] [PubMed] [Google Scholar]
  22. Nava E., Palmer R. M., Moncada S. Inhibition of nitric oxide synthesis in septic shock: how much is beneficial? Lancet. 1991 Dec 21;338(8782-8783):1555–1557. doi: 10.1016/0140-6736(91)92375-c. [DOI] [PubMed] [Google Scholar]
  23. Nunez D. J., Dickson M. C., Brown M. J. Natriuretic peptide receptor mRNAs in the rat and human heart. J Clin Invest. 1992 Nov;90(5):1966–1971. doi: 10.1172/JCI116075. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Nunokawa Y., Ishida N., Tanaka S. Cloning of inducible nitric oxide synthase in rat vascular smooth muscle cells. Biochem Biophys Res Commun. 1993 Feb 26;191(1):89–94. doi: 10.1006/bbrc.1993.1188. [DOI] [PubMed] [Google Scholar]
  25. Petros A., Bennett D., Vallance P. Effect of nitric oxide synthase inhibitors on hypotension in patients with septic shock. Lancet. 1991 Dec 21;338(8782-8783):1557–1558. doi: 10.1016/0140-6736(91)92376-d. [DOI] [PubMed] [Google Scholar]
  26. Riveros-Moreno V., Beddell C., Moncada S. Nitric oxide synthase. Structural studies using anti-peptide antibodies. Eur J Biochem. 1993 Aug 1;215(3):801–808. doi: 10.1111/j.1432-1033.1993.tb18095.x. [DOI] [PubMed] [Google Scholar]
  27. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Scott-Burden T., Schini V. B., Elizondo E., Junquero D. C., Vanhoutte P. M. Platelet-derived growth factor suppresses and fibroblast growth factor enhances cytokine-induced production of nitric oxide by cultured smooth muscle cells. Effects on cell proliferation. Circ Res. 1992 Nov;71(5):1088–1100. doi: 10.1161/01.res.71.5.1088. [DOI] [PubMed] [Google Scholar]
  29. Severn A., Wakelam M. J., Liew F. Y. The role of protein kinase C in the induction of nitric oxide synthesis by murine macrophages. Biochem Biophys Res Commun. 1992 Nov 16;188(3):997–1002. doi: 10.1016/0006-291x(92)91330-s. [DOI] [PubMed] [Google Scholar]
  30. Szabó C., Mitchell J. A., Gross S. S., Thiemermann C., Vane J. R. Nifedipine inhibits the induction of nitric oxide synthase by bacterial lipopolysaccharide. J Pharmacol Exp Ther. 1993 May;265(2):674–680. [PubMed] [Google Scholar]
  31. Xie Q. W., Whisnant R., Nathan C. Promoter of the mouse gene encoding calcium-independent nitric oxide synthase confers inducibility by interferon gamma and bacterial lipopolysaccharide. J Exp Med. 1993 Jun 1;177(6):1779–1784. doi: 10.1084/jem.177.6.1779. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Yamamoto K. K., Gonzalez G. A., Biggs W. H., 3rd, Montminy M. R. Phosphorylation-induced binding and transcriptional efficacy of nuclear factor CREB. Nature. 1988 Aug 11;334(6182):494–498. doi: 10.1038/334494a0. [DOI] [PubMed] [Google Scholar]
  33. Zembowicz A., Vane J. R. Induction of nitric oxide synthase activity by toxic shock syndrome toxin 1 in a macrophage-monocyte cell line. Proc Natl Acad Sci U S A. 1992 Mar 15;89(6):2051–2055. doi: 10.1073/pnas.89.6.2051. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from British Journal of Pharmacology are provided here courtesy of The British Pharmacological Society

RESOURCES