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. 2001 May;56(5):351–357. doi: 10.1136/thorax.56.5.351

Increased expression of inducible nitric oxide synthase and cyclo-oxygenase-2 in the airway epithelium of asthmatic subjects and regulation by corticosteroid treatment

A Redington 1, Q Meng 1, D Springall 1, T Evans 1, C Creminon 1, J Maclouf 1, S Holgate 1, P Howarth 1, J Polak 1
PMCID: PMC1746058  PMID: 11312402

Abstract

BACKGROUND—Nitric oxide (NO) and prostanoids are mediators of vascular and bronchial tone that are postulated to be involved in asthma. Increased levels of both are found in asthmatic subjects and are synthesised by enzymes that have cytokine inducible forms: inducible NO synthase (iNOS) and cyclo-oxygenase-2 (COX-2), respectively. We hypothesised that the in vivo expression of iNOS and COX-2 in the airways would be increased in asthma, and that these cytokine inducible enzymes may represent targets for regulation by corticosteroid treatment.
METHODS—Bronchial biopsy specimens were obtained from three groups of subjects: atopic asthmatics treated with β2 agonists alone (n=7), atopic asthmatics additionally receiving regular treatment with corticosteroids (n=8), and non-asthmatic control subjects (n=10). Expression of iNOS and COX-2 mRNA and immunoreactive protein was studied using in situ hybridisation and quantitative immunohistochemistry.
RESULTS—Immunoreactivity and the hybridisation signal for iNOS and COX-2 were mainly localised in the airway epithelium. The proportion of epithelium immunostained was significantly greater in the non-steroid treated asthmatic subjects (iNOS 8.6 (1.8)%; COX-2 26.3 (4.6)%) than either the steroid treated asthmatics (iNOS 3.4 (1.0)%, p=0.009; COX-2 13.0 (0.6)%, p=0.0015) or the non-asthmatic controls (iNOS 4.2 (0.9)%, p=0.018; COX-2 11.6 (0.6)%, p=0.0003). Similarly, the hybridisation signal was stronger in the non-steroid treated group of asthmatic subjects than in the other two groups.
CONCLUSIONS—These findings highlight the potential role of the airway epithelium both as a contributor to the inflammatory process in asthma and as a target for inhaled corticosteroid treatment in this disease.



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Selected References

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  1. Alving K., Fornhem C., Lundberg J. M. Pulmonary effects of endogenous and exogenous nitric oxide in the pig: relation to cigarette smoke inhalation. Br J Pharmacol. 1993 Oct;110(2):739–746. doi: 10.1111/j.1476-5381.1993.tb13874.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Alving K., Weitzberg E., Lundberg J. M. Increased amount of nitric oxide in exhaled air of asthmatics. Eur Respir J. 1993 Oct;6(9):1368–1370. [PubMed] [Google Scholar]
  3. Campbell A. M., Chanez P., Vignola A. M., Bousquet J., Couret I., Michel F. B., Godard P. Functional characteristics of bronchial epithelium obtained by brushing from asthmatic and normal subjects. Am Rev Respir Dis. 1993 Mar;147(3):529–534. doi: 10.1164/ajrccm/147.3.529. [DOI] [PubMed] [Google Scholar]
  4. Churchill L., Chilton F. H., Resau J. H., Bascom R., Hubbard W. C., Proud D. Cyclooxygenase metabolism of endogenous arachidonic acid by cultured human tracheal epithelial cells. Am Rev Respir Dis. 1989 Aug;140(2):449–459. doi: 10.1164/ajrccm/140.2.449. [DOI] [PubMed] [Google Scholar]
  5. Coleman R. A., Sheldrick R. L. Prostanoid-induced contraction of human bronchial smooth muscle is mediated by TP-receptors. Br J Pharmacol. 1989 Mar;96(3):688–692. doi: 10.1111/j.1476-5381.1989.tb11869.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Demoly P., Jaffuel D., Lequeux N., Weksler B., Créminon C., Michel F. B., Godard P., Bousquet J. Prostaglandin H synthase 1 and 2 immunoreactivities in the bronchial mucosa of asthmatics. Am J Respir Crit Care Med. 1997 Feb;155(2):670–675. doi: 10.1164/ajrccm.155.2.9032211. [DOI] [PubMed] [Google Scholar]
  7. Evans T. J., Buttery L. D., Carpenter A., Springall D. R., Polak J. M., Cohen J. Cytokine-treated human neutrophils contain inducible nitric oxide synthase that produces nitration of ingested bacteria. Proc Natl Acad Sci U S A. 1996 Sep 3;93(18):9553–9558. doi: 10.1073/pnas.93.18.9553. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gustafsson L. E., Leone A. M., Persson M. G., Wiklund N. P., Moncada S. Endogenous nitric oxide is present in the exhaled air of rabbits, guinea pigs and humans. Biochem Biophys Res Commun. 1991 Dec 16;181(2):852–857. doi: 10.1016/0006-291x(91)91268-h. [DOI] [PubMed] [Google Scholar]
  9. Habib A., Créminon C., Frobert Y., Grassi J., Pradelles P., Maclouf J. Demonstration of an inducible cyclooxygenase in human endothelial cells using antibodies raised against the carboxyl-terminal region of the cyclooxygenase-2. J Biol Chem. 1993 Nov 5;268(31):23448–23454. [PubMed] [Google Scholar]
  10. Hamid Q., Springall D. R., Riveros-Moreno V., Chanez P., Howarth P., Redington A., Bousquet J., Godard P., Holgate S., Polak J. M. Induction of nitric oxide synthase in asthma. Lancet. 1993 Dec 18;342(8886-8887):1510–1513. doi: 10.1016/s0140-6736(05)80083-2. [DOI] [PubMed] [Google Scholar]
  11. Hardy C. C., Robinson C., Tattersfield A. E., Holgate S. T. The bronchoconstrictor effect of inhaled prostaglandin D2 in normal and asthmatic men. N Engl J Med. 1984 Jul 26;311(4):209–213. doi: 10.1056/NEJM198407263110401. [DOI] [PubMed] [Google Scholar]
  12. Heiss L. N., Lancaster J. R., Jr, Corbett J. A., Goldman W. E. Epithelial autotoxicity of nitric oxide: role in the respiratory cytopathology of pertussis. Proc Natl Acad Sci U S A. 1994 Jan 4;91(1):267–270. doi: 10.1073/pnas.91.1.267. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. Högman M., Frostell C. G., Hedenström H., Hedenstierna G. Inhalation of nitric oxide modulates adult human bronchial tone. Am Rev Respir Dis. 1993 Dec;148(6 Pt 1):1474–1478. doi: 10.1164/ajrccm/148.6_Pt_1.1474. [DOI] [PubMed] [Google Scholar]
  15. Kacmarek R. M., Ripple R., Cockrill B. A., Bloch K. J., Zapol W. M., Johnson D. C. Inhaled nitric oxide. A bronchodilator in mild asthmatics with methacholine-induced bronchospasm. Am J Respir Crit Care Med. 1996 Jan;153(1):128–135. doi: 10.1164/ajrccm.153.1.8542105. [DOI] [PubMed] [Google Scholar]
  16. Kharitonov S. A., Yates D. H., Barnes P. J. Inhaled glucocorticoids decrease nitric oxide in exhaled air of asthmatic patients. Am J Respir Crit Care Med. 1996 Jan;153(1):454–457. doi: 10.1164/ajrccm.153.1.8542158. [DOI] [PubMed] [Google Scholar]
  17. Kharitonov S. A., Yates D., Robbins R. A., Logan-Sinclair R., Shinebourne E. A., Barnes P. J. Increased nitric oxide in exhaled air of asthmatic patients. Lancet. 1994 Jan 15;343(8890):133–135. doi: 10.1016/s0140-6736(94)90931-8. [DOI] [PubMed] [Google Scholar]
  18. Kobzik L., Bredt D. S., Lowenstein C. J., Drazen J., Gaston B., Sugarbaker D., Stamler J. S. Nitric oxide synthase in human and rat lung: immunocytochemical and histochemical localization. Am J Respir Cell Mol Biol. 1993 Oct;9(4):371–377. doi: 10.1165/ajrcmb/9.4.371. [DOI] [PubMed] [Google Scholar]
  19. Liu M. C., Bleecker E. R., Lichtenstein L. M., Kagey-Sobotka A., Niv Y., McLemore T. L., Permutt S., Proud D., Hubbard W. C. Evidence for elevated levels of histamine, prostaglandin D2, and other bronchoconstricting prostaglandins in the airways of subjects with mild asthma. Am Rev Respir Dis. 1990 Jul;142(1):126–132. doi: 10.1164/ajrccm/142.1.126. [DOI] [PubMed] [Google Scholar]
  20. Maier J. A., Hla T., Maciag T. Cyclooxygenase is an immediate-early gene induced by interleukin-1 in human endothelial cells. J Biol Chem. 1990 Jul 5;265(19):10805–10808. [PubMed] [Google Scholar]
  21. Mitchell J. A., Belvisi M. G., Akarasereenont P., Robbins R. A., Kwon O. J., Croxtall J., Barnes P. J., Vane J. R. Induction of cyclo-oxygenase-2 by cytokines in human pulmonary epithelial cells: regulation by dexamethasone. Br J Pharmacol. 1994 Nov;113(3):1008–1014. doi: 10.1111/j.1476-5381.1994.tb17093.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Newton R., Kuitert L. M., Slater D. M., Adcock I. M., Barnes P. J. Cytokine induction of cytosolic phospholipase A2 and cyclooxygenase-2 mRNA is suppressed by glucocorticoids in human epithelial cells. Life Sci. 1997;60(1):67–78. doi: 10.1016/s0024-3205(96)00590-5. [DOI] [PubMed] [Google Scholar]
  23. O'Banion M. K., Winn V. D., Young D. A. cDNA cloning and functional activity of a glucocorticoid-regulated inflammatory cyclooxygenase. Proc Natl Acad Sci U S A. 1992 Jun 1;89(11):4888–4892. doi: 10.1073/pnas.89.11.4888. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. O'Neill G. P., Ford-Hutchinson A. W. Expression of mRNA for cyclooxygenase-1 and cyclooxygenase-2 in human tissues. FEBS Lett. 1993 Sep 13;330(2):156–160. doi: 10.1016/0014-5793(93)80263-t. [DOI] [PubMed] [Google Scholar]
  25. Pang L., Pitt A., Petkova D., Knox A. J. The COX-1/COX-2 balance in asthma. Clin Exp Allergy. 1998 Sep;28(9):1050–1058. doi: 10.1046/j.1365-2222.1998.00311.x. [DOI] [PubMed] [Google Scholar]
  26. Persson M. G., Zetterström O., Agrenius V., Ihre E., Gustafsson L. E. Single-breath nitric oxide measurements in asthmatic patients and smokers. Lancet. 1994 Jan 15;343(8890):146–147. doi: 10.1016/s0140-6736(94)90935-0. [DOI] [PubMed] [Google Scholar]
  27. Redington A. E., Bradding P., Holgate S. T. The role of cytokines in the pathogenesis of allergic asthma. Reg Immunol. 1993 May-Aug;5(3-4):174–200. [PubMed] [Google Scholar]
  28. Redington A. E., Springall D. R., Ghatei M. A., Madden J., Bloom S. R., Frew A. J., Polak J. M., Holgate S. T., Howarth P. H. Airway endothelin levels in asthma: influence of endobronchial allergen challenge and maintenance corticosteroid therapy. Eur Respir J. 1997 May;10(5):1026–1032. doi: 10.1183/09031936.97.10051026. [DOI] [PubMed] [Google Scholar]
  29. Redington A. E., Springall D. R., Meng Q. H., Tuck A. B., Holgate S. T., Polak J. M., Howarth P. H. Immunoreactive endothelin in bronchial biopsy specimens: increased expression in asthma and modulation by corticosteroid therapy. J Allergy Clin Immunol. 1997 Oct;100(4):544–552. doi: 10.1016/s0091-6749(97)70148-9. [DOI] [PubMed] [Google Scholar]
  30. Ricciardolo F. L., Geppetti P., Mistretta A., Nadel J. A., Sapienza M. A., Bellofiore S., Di Maria G. U. Randomised double-blind placebo-controlled study of the effect of inhibition of nitric oxide synthesis in bradykinin-induced asthma. Lancet. 1996 Aug 10;348(9024):374–377. doi: 10.1016/s0140-6736(96)04450-9. [DOI] [PubMed] [Google Scholar]
  31. Robbins R. A., Barnes P. J., Springall D. R., Warren J. B., Kwon O. J., Buttery L. D., Wilson A. J., Geller D. A., Polak J. M. Expression of inducible nitric oxide in human lung epithelial cells. Biochem Biophys Res Commun. 1994 Aug 30;203(1):209–218. doi: 10.1006/bbrc.1994.2169. [DOI] [PubMed] [Google Scholar]
  32. Saleh D., Barnes P. J., Giaid A. Increased production of the potent oxidant peroxynitrite in the lungs of patients with idiopathic pulmonary fibrosis. Am J Respir Crit Care Med. 1997 May;155(5):1763–1769. doi: 10.1164/ajrccm.155.5.9154889. [DOI] [PubMed] [Google Scholar]
  33. Sarih M., Souvannavong V., Adam A. Nitric oxide synthase induces macrophage death by apoptosis. Biochem Biophys Res Commun. 1993 Mar 15;191(2):503–508. doi: 10.1006/bbrc.1993.1246. [DOI] [PubMed] [Google Scholar]
  34. Sherman P. A., Laubach V. E., Reep B. R., Wood E. R. Purification and cDNA sequence of an inducible nitric oxide synthase from a human tumor cell line. Biochemistry. 1993 Nov 2;32(43):11600–11605. doi: 10.1021/bi00094a017. [DOI] [PubMed] [Google Scholar]
  35. Sousa A. r., Pfister R., Christie P. E., Lane S. J., Nasser S. M., Schmitz-Schumann M., Lee T. H. Enhanced expression of cyclo-oxygenase isoenzyme 2 (COX-2) in asthmatic airways and its cellular distribution in aspirin-sensitive asthma. Thorax. 1997 Nov;52(11):940–945. doi: 10.1136/thx.52.11.940. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Springall D. R., Riveros-Moreno V., Buttery L., Suburo A., Bishop A. E., Merrett M., Moncada S., Polak J. M. Immunological detection of nitric oxide synthase(s) in human tissues using heterologous antibodies suggesting different isoforms. Histochemistry. 1992 Nov;98(4):259–266. doi: 10.1007/BF00271040. [DOI] [PubMed] [Google Scholar]
  37. Taha R., Olivenstein R., Utsumi T., Ernst P., Barnes P. J., Rodger I. W., Giaid A. Prostaglandin H synthase 2 expression in airway cells from patients with asthma and chronic obstructive pulmonary disease. Am J Respir Crit Care Med. 2000 Feb;161(2 Pt 1):636–640. doi: 10.1164/ajrccm.161.2.9811063. [DOI] [PubMed] [Google Scholar]
  38. Walenga R. W., Kester M., Coroneos E., Butcher S., Dwivedi R., Statt C. Constitutive expression of prostaglandin endoperoxide G/H synthetase (PGHS)-2 but not PGHS-1 in hum an tracheal epithelial cells in vitro. Prostaglandins. 1996 Nov;52(5):341–359. doi: 10.1016/s0090-6980(96)00101-3. [DOI] [PubMed] [Google Scholar]
  39. Watkins D. N., Peroni D. J., Lenzo J. C., Knight D. A., Garlepp M. J., Thompson P. J. Expression and localization of COX-2 in human airways and cultured airway epithelial cells. Eur Respir J. 1999 May;13(5):999–1007. doi: 10.1034/j.1399-3003.1999.13e12.x. [DOI] [PubMed] [Google Scholar]
  40. Yates D. H., Kharitonov S. A., Robbins R. A., Thomas P. S., Barnes P. J. Effect of a nitric oxide synthase inhibitor and a glucocorticosteroid on exhaled nitric oxide. Am J Respir Crit Care Med. 1995 Sep;152(3):892–896. doi: 10.1164/ajrccm.152.3.7663801. [DOI] [PubMed] [Google Scholar]
  41. Yokoyama C., Tanabe T. Cloning of human gene encoding prostaglandin endoperoxide synthase and primary structure of the enzyme. Biochem Biophys Res Commun. 1989 Dec 15;165(2):888–894. doi: 10.1016/s0006-291x(89)80049-x. [DOI] [PubMed] [Google Scholar]

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