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Arzneimittelforschung 2011; 61(12): 700-706
DOI: 10.1055/s-0031-1300590
Antioxidants
Editio Cantor Verlag Aulendorf (Germany)

Protective effect of erdosteine metabolite I against hydrogen peroxide-induced oxidative DNA-damage in lung epithelial cells

Laura Marabini
1   Department of Pharmacology, Chemotherapy and Medical Toxicology, School of Medicine, University of Milan, Milan, Italy
,
Rossella Calò
1   Department of Pharmacology, Chemotherapy and Medical Toxicology, School of Medicine, University of Milan, Milan, Italy
,
Pier Carlo Braga
1   Department of Pharmacology, Chemotherapy and Medical Toxicology, School of Medicine, University of Milan, Milan, Italy
› Author Affiliations
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Publication History

Publication Date:
09 February 2012 (online)

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Abstract

It has been shown that the mucolytic agent erdosteine (N-carboxymethylthio-acetyl-homocysteine thiolactone, CAS 84611–23–4) has anti-inflammatory and anti-oxidant properties, and an active metabolite I (MET I) containing pharmacologically active sulphydryl group has been found to have a free radical scavenging activity.

The aim of this study was to assess the ability of erdosteine metabolite I to protect A549 human lung adenocarcinoma cell against hydrogen peroxide (H2O2)-mediated oxidative stress and oxidative DNA damage.

When A549 cells were pre-treated with the active metabolite I (2.5–5–10 µg/ml) for 10–30 min and then exposed to H2O2 (1–4 mM) for two additional hours at 37 °C, 5% at CO2, the intracellular peroxide production, reflected by dichloro-fluorescein (DCF) fluorescence, decreased in a concentration-dependent manner. Furthermore, using a comet assay as an indicator for oxidative DNA damage, it was found that the metabolite I prevented damage to cells exposed to short-term H2O2 treatment.

The data suggest that this compound is effective in preventing H2O2-induced oxidative stress and DNA damage in A549 cells. The underlying mechanisms involve the scavenging of intracellular reactive oxygen species (ROS).

Key words

A549 cells - Anti-oxidant activity - DNA damage - Erdosteine metabolite I - Genotoxicity
 

  • References

  • 1 MacNee W. Oxidants/antioxidants and COPD. Chest. 2000; 117 (Suppl 1) 303S-317S
    CrossrefPubMedGoogle Scholar
  • 2 Tao F, Gonzales-Flecha B, Kobzik L. Reactive oxygen species in pulmonary inflammation by ambient particulates. Free Radic Biol Med. 2003; 35: 327-40
    CrossrefPubMedGoogle Scholar
  • 3 Rahman I. Oxidative stress, transcription factors and chromatic remodelling in lung inflammation. Biochem Pharmacol. 2002; 64: 935-42
    CrossrefPubMedGoogle Scholar
  • 4 Fuschillo S, De Felice A, Balzano A. Mucosal inflammation in idiopathic bronchiectasis: cellular and molecular mechanisms. Eur Resp J. 2008; 31: 396-406
    CrossrefPubMedGoogle Scholar
  • 5 Barnes PJ. Reactive oxygen species and airway inflammation. Free Rad Biol Med. 1990; 9: 235-43
    CrossrefPubMedGoogle Scholar
  • 6 Dickinson DA, Forman HJ. Cellular glutathione and thiolmetabolism. Biochem Pharmacol. 2002; 64: 1019-26
    CrossrefPubMedGoogle Scholar
  • 7 Forman HJ, Liu RM, Shi MM. Glutathione synthesis in oxidative stress. In: Packer L, Cadenas E. editors Biothiols in health and diseases. New York: Marcel Dekker; 1995. p. 189-212
    Google Scholar
  • 8 Cantin AM, North SL, Hubbard RC, Crystal RG. Normal alveolar epithelial lining fluid contains high levels of glutathione. J Appl Physiol. 1987; 63: 152-7
    PubMedGoogle Scholar
  • 9 Patterson CE, Rhoades RA. Protective role of sulphydryl reagents in oxidant lung injury. Exp Lung Res. 1988; 14: 1005-19
    CrossrefPubMedGoogle Scholar
  • 10 Heffner JE, Repine JE. Pulmonary strategies of antioxidant defense. Am Rev Respir Dis. 1989; 140: 531-54
    CrossrefPubMedGoogle Scholar
  • 11 Rahman I. Regulation of glutathione in inflammation and chronic lung diseases. Mutat Res. 2005; 579: 58-80
    PubMedGoogle Scholar
  • 12 Braga PC, Dal Sasso M. Zuccotti. Assessment of the antioxidant activity of the SH-metabolite I of erdosteine on human neutrophil oxidative bursts. Arzneimittelforschung. 2000; 50: 739-46
    Article in Thieme ConnectPubMedGoogle Scholar
  • 13 Braga PC, Ziment I, Allegra L. Classification of agents that act on bronchial mucus. In: Braga PC, Allegra L. editors. Drugs in bronchial mucology. New York: Raven Press; 1989. p. 59-67
    Google Scholar
  • 14 Allegra L, Dal Sasso M, Bovio C, Massoni C, Fonti E, Braga PC. Human neutrophil oxidative bursts and their in vitro modulation by different N-acetylcysteine concentrations. Arzneimittelforschung. 2002; 52: 669-76
    Article in Thieme ConnectPubMedGoogle Scholar
  • 15 Braga PC. Oxidative stress: respiratory diseases and thiol compounds. Pisa (Italy): Pacini Editore; 2006. p. 58-63
    Google Scholar
  • 16 Dechant KL, Noble S. Erdosteine. Drugs. 1996; 52: 875-82
    CrossrefPubMedGoogle Scholar
  • 17 Dal Sasso M, Culici M, Bianchi T, Fonti E, Braga PC. Inhibitory effects of metabolite I of erdosteine on the generation of nitric oxide and peroxynitrite chemiluminescence by human neutrophils. Pharmacology. 2004; 71: 120-7
    CrossrefPubMedGoogle Scholar
  • 18 Dal Sasso M, Culici M, Guffanti EE, Bianchi T, Fonti E, Braga PC. A combination of budesonide and the SH-metabolite of erdosteine acts synergistically in reducing the chemiluminescence during human neutrophil respiratory burst. Pharmacology. 2005; 74: 127-34
    PubMedGoogle Scholar
  • 19 Braga PC, Dal Sasso M, Culici M, Verducci P, Lo Verso R, Marabini L. Effect of metabolite I of erdosteine on the release of human neutrophil elastase. Pharmacology. 2006; 77: 150-4
    CrossrefPubMedGoogle Scholar
  • 20 Braga PC, Culici M, Dal Sasso M, Falchi M, Spallino A. Free radical scavenging activity of erdosteine metabolite I investigated by electron paramagnetic resonance spectroscopy. Pharmacology. 2010; 85: 195-202
    CrossrefPubMedGoogle Scholar
  • 21 Foldbjerg R, Dang DA, Autrup H. Cytotoxicity and genotoxicity of silver nanoparticles in the human lung cancer cell line, A549. Arch Toxicol. 2010; DOI:10.1007/s00204-010-0545.
    PubMedGoogle Scholar
  • 22 Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951; 193: 265-75
    PubMedGoogle Scholar
  • 23 Singh NP, McCoy MT, Tice RR, Schneider EL. A simple technique for quantitation of low levels of DNA damage in individual cells. Exp Cell Res. 1988; 175: 184-91
    CrossrefPubMedGoogle Scholar
  • 24 Gminski R, Decker K, Heinz C, Seidel A, Könczöl M, Goldenberg E, et al. Genotoxic effects of three selected black toner powders and their dimethyl sulfoxide extracts in cultured human epithelial A549 lung cells in vitro. Environ Mol Mutagen. 2010; DOI:10.1002/em.20621.
    PubMed
  • 25 Di Simplicio P, Franconi F, Frosali S, Di Giuseppe D. Thio-lation and nitrosation of cysteines in biological fluids and cells. Amino Acids. 2003; 25: 323-39
    CrossrefPubMedGoogle Scholar
  • 26 Heins A, Mc Phail DB, Sokolowski T, Stockmann H, Schwarz K. The location of phenolic antioxidants and radicals at interfaces determines their activity. Lipids. 2007; 42: 573-82
    CrossrefPubMedGoogle Scholar
  • 27 Braga PC, Sambataro G, Dal Sasso M, Culici M, Alfieri M, Nappi G. Antioxidant effect of sulphurus thermal water on human neutrophil respiratory bursts: chemiluminescence evaluation. Respiration. 2008; 75: 193-204
    CrossrefPubMedGoogle Scholar
  • 28 Rahman I, Kilty I. Antioxidant therapeutic targets in COPD. Curr Drug Targets. 2006; 7: 707-20
    CrossrefPubMedGoogle Scholar
  • 29 Dal Negro RW, Visconti M, Micheletto C, Tomella S. Antioxidant effects of erdosteine in current smokers with mild COPD. Eur Respir J. 2006; 28 (Suppl 50) 429s
    PubMedGoogle Scholar
  • 30 Weissler JC. Idiopathic pulmonary fibrosis: cellular and molecular pathogenesis. Am J Med Sci. 1989; 297: 91-104
    CrossrefPubMedGoogle Scholar
  • 31 Paredi P, Kharitonov SA, Loukides S, Pantelidis P, du Bois RM, Barnes PJ. Exhaled nitric oxide is increased in active fibrosing alveolitis. Chest. 1999; 115: 1352-6
    CrossrefPubMedGoogle Scholar
  • 32 Basyigit I, Yildiz F, Cekmen M, Duman C, Bulut O. Effects of erdosteine on smoking-induced lipid peroxidation in healty smokers. Drugs R D. 2005; 6: 83-9
    CrossrefPubMedGoogle Scholar
  • 33 Moretti M, Marchioni CF. An overview of erdosteine antioxidant activity in experimental research. Pharmacol Res. 2007; 55: 249-54
    CrossrefPubMedGoogle Scholar
  • 34 Morley N, Curnow A, Salter S, Campbell S, Gould D. N-acetyl-L-cysteine prevents DNA damage induced by UVA, UVB and visible radiation in human fibroblasts. J Photo-chem Photobiol. 2003; 72: 55-60
    CrossrefPubMedGoogle Scholar
  • 35 Salter L, Clifford T, Morley N, Gould D, Campbell S, Curnow A. The use of comet assay data with a simple reaction mechanism to evaluate the relative effectiveness of free radical scavenging by quercitin, epigallocatechin gallate and N-acetylcysteine in UV-irradiated MRC5 lung fibroblasts. J Photochem Photobiol. 2004; 75: 57-61
    CrossrefPubMedGoogle Scholar
  • 36 Tiwari P, Kumar A, Balakrishnam S, Kushwara HS, Mishra KP. Radiation-induced micronucleus formation and DNA damage in human lymphocytes and their prevention by antioxidant thiols. Mutat Res. 2009; 676 (l–2) 62-8
    CrossrefPubMedGoogle Scholar
  • 37 Savu S, Mitrea M, Silvestro L, Mancini C. HPLC with online mass spectrometry detection application to elucidate erdosteine metabolism: preliminary study. Int J Clin Pharmacol. 2000; 38: 415-7
    PubMedGoogle Scholar