Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress
Clinical Collections
Advances in Alzheimer’s

Keynote webinar | Spotlight on medication adherence

LIVE: Thursday 27th June 2024, 18:00-19:30 (CEST)
Join our expert panel to discover why you need to understand the drivers of non-adherence in your patients, and how you can optimize medication adherence in your clinics to drastically improve patient outcomes.
ADVANCED SEARCH
Log in
Top
Respiratory Research
Published in: Respiratory Research 1/2005

Open Access 01-12-2005 | Research

Inhibition or knock out of Inducible nitric oxide synthase result in resistance to bleomycin-induced lung injury

Authors: Tiziana Genovese, Salvatore Cuzzocrea, Rosanna Di Paola, Marco Failla, Emanuela Mazzon, Maria Angela Sortino, Giuseppina Frasca, Elisa Gili, Nunzio Crimi, Achille P Caputi, Carlo Vancheri

Published in: Respiratory Research | Issue 1/2005

Login to get access

Abstract

Background

In the present study, by comparing the responses in wild-type mice (WT) and mice lacking (KO) the inducible (or type 2) nitric oxide synthase (iNOS), we investigated the role played by iNOS in the development of on the lung injury caused by bleomycin administration. When compared to bleomycin-treated iNOSWT mice, iNOSKO mice, which had received bleomycin, exhibited a reduced degree of the (i) lost of body weight, (ii) mortality rate, (iii) infiltration of the lung with polymorphonuclear neutrophils (MPO activity), (iv) edema formation, (v) histological evidence of lung injury, (vi) lung collagen deposition and (vii) lung Transforming Growth Factor beta1 (TGF-β1) expression.

Methods

Mice subjected to intratracheal administration of bleomycin developed a significant lung injury. Immunohistochemical analysis for nitrotyrosine revealed a positive staining in lungs from bleomycin-treated iNOSWT mice.

Results

The intensity and degree of nitrotyrosine staining was markedly reduced in tissue section from bleomycin-iNOSKO mice. Treatment of iNOSWT mice with of GW274150, a novel, potent and selective inhibitor of iNOS activity (5 mg/kg i.p.) also significantly attenuated all of the above indicators of lung damage and inflammation.

Conclusion

Taken together, our results clearly demonstrate that iNOS plays an important role in the lung injury induced by bleomycin in the mice.
Literature
1.
2.
Crouch E: Pathobiology of pulmonary fibrosis. Am J Physiol 1990, 259:L159–84.PubMed
3.
Crystal RG, Bitterman PB, Rennard SI, Hance AJ, Keogh BA: Interstitial lung diseases of unknown cause: disorders characterized by chronic inflammation of the lower respiratory tract. N Engl J Med 1984, 310:154–66.CrossRefPubMed
4.
5.
Nathan C: Nitric oxide as a secretary product of mammalian cells. FASEB J 1992, 6:3051–3064.PubMed
6.
Dinerman J: Molecular mechanism of nitric oxide production. Potential relevance to cardiovascolar disease. Circ Res 1993, 73:217–222.CrossRefPubMed
7.
Szabò C: Alterations in the production of NO in various forms of circulatory shock. New Horiz 1995, 3:3–32.
8.
Southan G, Szabò C: Selective pharmacological inhibition of distinct NOS isoforms. Bioch Pharm 1996, 51:383–394.CrossRef
9.
Moncada S, Palmer RMJ: NO: physiology, patophysiology and pharmacology. Pharm Rev 1991, 43:109–142.PubMed
10.
Moncada S, Higgs A: The L-arginine NO pathway. New Eng J of Med 2002, 329:2002–2012.
11.
Cuzzocrea S: Effect of inhibitors of nitric oxide in animal models and future directions for therapy in inflammatory disorders. Current Med Chem Anti-inflammatory & Anti-allergy agent 2004, 3:261–270.CrossRef
12.
Crow JP, Beckman JS: Reactions between nitric oxide, superoxide, and peroxynitrite: footprints of peroxynitrite in vivo. Adv Pharmacol 1995, 34:17–43.CrossRefPubMed
13.
Pryor WA, Squadrito GL: The chemistry of peroxinitrite: a product from reaction of NO with superoxide. Am J Physiol 1995, 268:699–722.
14.
Beckman JS, Beckman TW, Chen J, Marshall PA, Freeman BA: Apparent hydroxil radical production by peroxynitrite: implications for endothelial injury from NO and superoxide. Proc Natl Acad Sci USA 1990, 87:1620–1624.CrossRefPubMedPubMedCentral
15.
Cuzzocrea S, Riley DP, Caputi AP, Salvemini D: Antioxidant therapy: a new pharmacological approach in shock, inflammation, and ischemia/reperfusion injury. Pharmacol Rev 2001, 53:135–159.PubMed
16.
Rao AV, Balachandran B: Role of oxidative stress and antioxidants in neurodegenerative diseases. Nutr Neurosci 2002, 5:291–309.CrossRefPubMed
17.
Iuliano L: The oxidant stress hypothesis of atherogenesis. Lipids 2001, 36:41–44.CrossRef
18.
Yamada T, Grisham MB: Role of neutrophil-derived oxidants in the pathogenesis of intestinal inflammation. Klin Wochenschr 1991, 69:988–994.CrossRefPubMed
19.
Inoue S, Kawanishi S: Oxidative DNA damage induced by simultaneous generation of nitric oxide and superoxide. FEBS Lett 1995, 371:86–88.CrossRefPubMed
20.
Salgo MG: Peroxinitrite causes apoptosis in rat thymorytes. Bioch Bioph Res Commun 1995, 215:1111–1118.CrossRef
21.
22.
23.
Young RJ, Beams RM, Carter K, Clark HA, Coe DM, Chambers CL, Davies PI, Dawson J, Drysdale MJ, Franzman KW, French C, Hodgson ST, Hodson HF, Kleanthous S, Rider P, Sanders D, Sawyer DA, Scott KJ, Shearer BG, Stocker R, Smith S, Tackley MC, Knowles RG: Inhibition of inducible nitric oxide synthase by acetamidine derivatives of hetero-substituted lysine and homolysine. Bioorg Med Chem Lett 2000, 10:597–600.CrossRefPubMed
24.
Mcdonald MC, Izumi M, Cuzzocrea S, Thiemermann C: A novel, potent and selective inhibitor of the activity of inducible nitric oxide synthase (GW274150) reduces the organ injury in hemorrhagic shock. J Physiol Pharmacol 2002, 53:555–569.PubMed
25.
Chatterjee PK, Patel NS, Sivarajah A, Kvale EO, Dugo L, Cuzzocrea S, Brown PA, Stewart KN, Mota-Filipe H, Britti D, et al.: GW274150, a potent and highly selective inhibitor of iNOS, reduces experimental renal ischemia/reperfusion injury. Kidney Int 2003, 63:853–865.CrossRefPubMed
26.
Cuzzocrea S, Chatterjee PK, Mazzon E, Mcdonald MC, Dugo L, Di Paola R, Serraino I, Britti D, Caputi AP, Thiemermann C: Beneficial effects of GW274150, a novel, potent and selective inhibitor of iNOS activity, in a rodent model of collagen-induced arthritis. Eur J Pharmacol 2002, 453:119–129.CrossRefPubMed
27.
Dugo L, Marzocco S, Mazzon E, Di Paola R, Genovese T, Caputi AP, Cuzzocrea S: Effects of GW274150, a novel and selective inhibitor of iNOS activity, in acute lung inflammation. Br J Pharmacol 2004,141(6):979–87.CrossRefPubMedPubMedCentral
28.
29.
Cuzzocrea S, Ianaro A, Wayman NS, Mazzon E, Pisano B, Dugo L, Serraino I, Di Paola R, Chatterjee PK, Di Rosa M, et al.: The cyclopentenone prostaglandin 15-deoxy-delta-(12,14)-PGJ2 attenuates the development of colon injury caused by dinitrobenzene sulphonic acid in the rat. Br J Pharmacol 2003, 138:678–88.CrossRefPubMedPubMedCentral
30.
Mullane KM, Kraemer R, Smith B: Myeloperoxidase activity as a quantitative assessment of neutrophil infiltration into ischemic myocardium. J Pharmacol Methods 1985, 14:157–67.CrossRefPubMed
31.
Bradford M: A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of proteindye binding. Anal Biochem 1976, 72:248–54.CrossRefPubMed
32.
33.
34.
Hardie WD, Le Cras TD, Jiang K, Tichelaar JW, Azhar M, Korfhagen TR: Conditional expression of transforming growth factor-alpha in adult mouse lung causes pulmonary fibrosis. Am J Physiol Lung Cell Mol Physiol 2004, 286:L741-L749.CrossRefPubMed
35.
Tang W, Geba GP, Zheng T, Ray P, Homer RJ, Kuhn C 3rd, Flavell RA, Elias JA: Targeted expression of IL-11 in the murine airway causes lymphocytic inflammation, bronchial remodeling, and airways obstruction. J Clin Invest 1996, 98:2845–53.CrossRefPubMedPubMedCentral
36.
Chandler DB, Hyde DM, Giri SN: Morphometric estimates of infiltrative cellular changes during the development of bleomycin-induced pulmonary fibrosis in hamsters. Am J Pathol 1983, 112:170–7.PubMedPubMedCentral
37.
38.
Burger RM, Projan SJ, Horwitz SB, Peisach J: The DNA cleavage mechanism of iron-bleomycin. Kinetic resolution of strand scission from base propenal release. J Biol Chem 1986, 261:15955–15959.PubMed
39.
Arslan SO, Zerin M, Vural H, Coskun A: The effect of melatonin on bleomycin-induced pulmonary fibrosis in rats. J Pineal Res 2002, 32:21–5.CrossRefPubMed
40.
Slosman DO, Costabella PM, Roth M, Werlen G, Polla BS: Bleomycin primes monocytes-macrophages for superoxide production. Eur Respir J 1990, 3:772–8.PubMed
41.
Goodman MT, Hernandez B, Wilkens LR, Lee J, Le Marchand L, Liu LQ, Franke AA, Kucuk O, Hsu TC: Effects of beta-carotene and alpha-tocopherol on bleomycin-induced chromosomal damage. Cancer Epidemiol Biomarkers Prev 1998, 7:113–7.PubMed
42.
Giri SN, Chen ZL, Younker WR, Schiedt MJ: Effects of intratracheal administration of bleomycin on GSH-shuttle enzymes, catalase, lipid peroxidation, and collagen content in the lungs of hamsters. Toxicol Appl Pharmacol 1983, 71:132–41.CrossRefPubMed
43.
Karam H, Hurbain-Kosmath I, HousSET B: Antioxidant activity in alveolar epithelial type 2 cells of rats during the development of bleomycin injury. Cell Biol Toxicol 1998, 14:13–22.CrossRefPubMed
44.
Ikezaki S, Nishikawa A, Enami T, Furukawa F, Imazawa T, Uneyama C, Fukushima S, TakahASHI M: Inhibitory effects of the dietary antioxidants butylated hydroxyanisole and butylated hydroxytoluene on bronchioloalveolar cell proliferation during the bleomycin-induced pulmonary fibrosing process in hamsters. Food Chem Toxicol 1996, 34:327–35.CrossRefPubMed
45.
Venkatesan N, Punithavathi V, Chandrakasan G: Curcumin protects bleomycin-induced lung injury in rats. Life Sci 1997, 61:PL51–8.CrossRefPubMed
46.
Cuzzocrea S, Zingarelli B, Gilard E, Hake P, Salzman AL, Szabó C: Anti-inflammatory effects of mercaptoethylguanidine, a combined inhibitor of nitric oxide synthase and peroxynitrite scavenger, in carrageenan-induced models of inflammation. Free Rad Biol Med 1998, 24:450–459.CrossRefPubMed
47.
Salvemini D, Wang ZQ, Wyatt P, Bourdon DM, Marino MH, Manning PT, Currie MG: Nitric oxide: a key mediator in the early and late phase of carrageenan-induced rat paw inflammation. Br J Pharmacol 1996, 118:829–838.CrossRefPubMedPubMedCentral
48.
Tracey WR, Nakane M, Kuk J, Budzik G, Klinghofer V, Harris R, Carter G: The nitric oxide synthase inhibitor, L-N G -monomethylarginine, reduces carrageenan-induced pleurisy in the rat. J Pharmacol Exp Ther 1995, 273:1295–1299.PubMed
49.
Wei XQ, Charles IG, Smith A, Ure J, Feng GJ, Huang FP, Xu D, Muller W, Moncada S, Liew FY: Altered immune responses in mice lacking inducible nitric oxide synthase. Nature 1995, 375:408–11.CrossRefPubMed
50.
El-Khatib AS: Possible modulatory role of nitric oxide in lung toxicity induced in rats by chronic administration of bleomycin. Chemotherap 2002, 48:244–51.CrossRef
51.
Jang AS, Lee JU, Choi IS, Park KO, Lee JH, Park SW, Park CS: Expression of nitric oxide synthase, aquaporin 1 and aquaporin 5 in rat after bleomycin inhalation. Intensive Care Med 2004, 30:489–95.CrossRefPubMed
52.
Davis DW, Weidner DA, Holian A, McConkey DJ: Nitric oxide-dependent activation of p53 suppresses bleomycin-induced apoptosis in the lung. J Exp Med 2000, 192:857–69.CrossRefPubMedPubMedCentral
53.
de Rezende MC, Martinez JA, Capelozzi VL, Simoes MJ, Beppu OS: Protective effect of aminoguanidine in a murine model of pulmonary fibrosis induced by bleomycin. Fundam Clin Pharmacol 2000, 14:561–7.CrossRefPubMed
54.
Hu J, Xu Q, Li B: The effect of aminoguanidine, a nitric oxide synthase inhibitor, on bleomycin-induced lung injury in rats. Zhonghua Jie He He Hu Xi Za Zhi 1999, 22:51–3.PubMed
55.
Yildirim Z, Turkoz Y, Kotuk M, Armutcu F, Gurel A, Iraz M, Ozen S, Aydogdu I, Akyol O: Effects of aminoguanidine and antioxidant erdosteine on bleomycin-induced lung fibrosis in rats. Nitric Oxide 2004, 11:156–65.CrossRefPubMed
56.
Giri SN, Biring I, Nguyen T, Wang Q, Hyde DM: Abrogation of bleomycin-induced lung fibrosis by nitric oxide synthase inhibitor, aminoguanidine in mice. Nitric Oxide 2002, 7:109–18.CrossRefPubMed
57.
Chen XL, Huang SS, Li WB, Wang DH, Wang XL: Inhibitory effect of aminoguanidine on bleomycin-induced pulmonary toxicity in rat. Acta Pharmacol Sin 2001, 22:711–5.PubMed
58.
59.
Bieganski T, Kusche J, Lorenz W, Hesterberg R, Stahlknecht CD, Feussner KD: Distribution and Properties of Human Intestinal Diamine Oxidase and Its Relevance for the Histamine Catabolism. Biochim Biophys Acta 1983, 756:196–203.CrossRefPubMed
60.
Nilsson BO, Kockum I, Rosengren E: Inhibition of Diamine Oxidase Promotes Uptake of Putrescine From Rat Small Intestine. Inflamm Res 1996, 45:513–518.CrossRefPubMed
61.
Ou P, Wolff SP: Aminoguanidine: a Drug Proposed for Prophylaxis in Diabetes Inhibits Catalase and Generates Hydrogen Peroxide in Vitro. Biochem Pharmacol 1993, 46:1139–1144.CrossRefPubMed
62.
Giardino I, Fard AK, Hatchell DL, Brownlee M: Aminoguanidine Inhibits Reactive Oxygen Species Formation, Lipid Peroxidation, and Oxidant-Induced Apoptosis. Diabetes 1998, 47:1114–1120.CrossRefPubMed
63.
Yildiz G, Demiryurek AT, Sahin-Erdemli I, Kanzik I: Comparison of Antioxidant Activities of Aminoguanidine, Methylguanidine and Guanidine by Luminol-Enhanced Chemiluminescence. Br J Pharmacol 1998, 124:905–910.CrossRefPubMedPubMedCentral
64.
Fine A, Goldstein RH: The effect of transforming growth factor on cell proliferation and collagen by fibroblasts. J Biol Chem 1987, 262:3897–3902.PubMed
65.
Sporn MB, Roberts AB, Wakefield LM, De Crombrugghe B: Some recent advances in the chemistry and biology of transforming growth factor-beta. J Cell Biol 1987, 105:1039–1045.CrossRefPubMed
66.
Pierce GF, Mustoe TA, Lingelbach J, Masakowski VR, Gramates P, Deuel TF: Transforming growth factor reverses the glucocorticoid-induced wound-healing deficit in rats: possible regulation in macrophages by platelet-derived growth factor. Proc Natl Acad Sci USA 1989, 86:2229–2233.CrossRefPubMedPubMedCentral
67.
Hoyt DG, Lazo JS: Alterations in pulmonary mRNA encoding procollagens, fibronectin and transforming growth factor precede bleomycin induced pulmonary fibrosis in mice. J Pharmacol Exp Ther 1988, 246:765–771.PubMed
68.
Raghow R, Irish P, Kang AH: Coordinate regulation of transforming growth factor gene expression and cell proliferation in hamster lungs undergoing bleomycin-induced pulmonary fibrosis. J Clin Invest 1989, 84:1836–1842.CrossRefPubMedPubMedCentral
69.
Khalil N, Bereznay O, Sporn M, Greenberg AH: Macrophage production of transforming growth factor and fibroblast collagen synthesis in chronic pulmonary inflammation. J Exp Med 1989, 170:727–737.CrossRefPubMed
70.
Khalil N, Corne S, Whitman C, Yacyshyn H: Plasmin regulates the activation of cell-associated latent TGF-1 secreted by rat alveolar macrophages after in vivo bleomycin injury. Am J Respir Cell Mol Biol 1996, 15:252–259.CrossRefPubMed
71.
Chatham WW, Swaim R, Frohsin H Jr, Heck LW, Miller EJ, Blackburn WD Jr: Degradation of human articular cartilage by neutrophils in synovial fluid. Arthritis Rheum 1993, 36:51–8.CrossRefPubMed
72.
Bowler RP, Nicks M, Warnick K, Crapo JD: Role of extracellular superoxide dismutase in bleomycin-induced pulmonary fibrosis. Am J Physiol Lung Cell Mol Physiol 2002, 282:L719–26.CrossRefPubMed
73.
Chen XL, Li WB, Zhou AM, Ai J, Huang SS: Role of endogenous peroxynitrite in pulmonary injury and fibrosis induced by bleomycin A5 in rats. Acta Pharmacol Sin 2003, 24:697–702.PubMed
74.
Beckman JS: Oxidative damage and tyrosine nitration from peroxynitrite. Chem Res Toxicol 1996, 9:836–44.CrossRefPubMed
75.
Eiserich JP, Hristova M, Cross CE, Jones AD, Freeman BA, Halliwell B, Van der Vliet A: Formation of nitric oxide-derived inflammatory oxidants by myeloperoxidase in neutrophils. Nature 1998, 391:393–397.CrossRefPubMed
Metadata
Title
Inhibition or knock out of Inducible nitric oxide synthase result in resistance to bleomycin-induced lung injury
Authors
Tiziana Genovese
Salvatore Cuzzocrea
Rosanna Di Paola
Marco Failla
Emanuela Mazzon
Maria Angela Sortino
Giuseppina Frasca
Elisa Gili
Nunzio Crimi
Achille P Caputi
Carlo Vancheri
Publication date
01-12-2005
Publisher
BioMed Central
Published in
Respiratory Research / Issue 1/2005
Electronic ISSN: 1465-993X
DOI
https://doi.org/10.1186/1465-9921-6-58

Other articles of this Issue 1/2005

Respiratory Research 1/2005 Go to the issue

Research

The absence of reactive oxygen species production protects mice against bleomycin-induced pulmonary fibrosis

Research

Expression profiling of laser-microdissected intrapulmonary arteries in hypoxia-induced pulmonary hypertension

Research

Activation of the SPHK/S1P signalling pathway is coupled to muscarinic receptor-dependent regulation of peripheral airways

Review

Anaphylatoxin C3a receptors in asthma

Research

IL-4 increases type 2, but not type 1, cytokine production in CD8+ T cells from mild atopic asthmatics

Research

Proinflammatory cytokine responses induced by influenza A (H5N1) viruses in primary human alveolar and bronchial epithelial cells
Live Webinar | 27-06-2024 | 18:00 (CEST)

Keynote webinar | Spotlight on medication adherence

Reserve your place

Live: Thursday 27th June 2024, 18:00-19:30 (CEST)

WHO estimates that half of all patients worldwide are non-adherent to their prescribed medication. The consequences of poor adherence can be catastrophic, on both the individual and population level.

Join our expert panel to discover why you need to understand the drivers of non-adherence in your patients, and how you can optimize medication adherence in your clinics to drastically improve patient outcomes.

Prof. Kevin Dolgin
Prof. Florian Limbourg
Prof. Anoop Chauhan
Developed by: Springer Medicine
Obesity Clinical Trial Summary

At a glance: The STEP trials

Read more

A round-up of the STEP phase 3 clinical trials evaluating semaglutide for weight loss in people with overweight or obesity.

Developed by: Springer Medicine

Highlights from the ACC 2024 Congress

Year in Review: Pediatric cardiology

Pediatric Cardiology Video

Watch Dr. Anne Marie Valente present the last year's highlights in pediatric and congenital heart disease in the official ACC.24 Year in Review session.

Year in Review: Pulmonary vascular disease

Cardiopulmonary Comorbidity Video

The last year's highlights in pulmonary vascular disease are presented by Dr. Jane Leopold in this official video from ACC.24.

Year in Review: Valvular heart disease

Valvular Heart Disease Video

Watch Prof. William Zoghbi present the last year's highlights in valvular heart disease from the official ACC.24 Year in Review session.

Year in Review: Heart failure and cardiomyopathies

Heart Failure Video

Watch this official video from ACC.24. Dr. Biykem Bozkurt discusses last year's major advances in heart failure and cardiomyopathies.

ACC 2024 Congress Coverage

Year in Review: Heart failure and cardiomyopathies

Heart Failure Video

Watch this official video from ACC.24. Dr. Biykem Bozkurt discusses last year's major advances in heart failure and cardiomyopathies.

Watch now

Semaglutide benefits people with type 2 diabetes and obesity-related heart failure

16-04-2024 Type 2 Diabetes News

Incentivised to exercise

11-04-2024 Lifestyle Modifications News

New intervention for STEMI-related cardiogenic shock

10-04-2024 Myocardial Infarction News

» View more highlights on the ACC 2024 congress hub page

Image Credits