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Thromb Haemost 2005; 94(02): 327-335
DOI: 10.1160/TH04-06-0360
Theme Issue Article
Schattauer GmbH

Chlamydia pneumoniae induces nitric oxide synthase and lipoxygenase- dependent production of reactive oxygen species in platelets

Effects on oxidation of low density lipoproteins
Hanna Kälvegren
1   Department of Medicine and Care, Division of Pharmacology
,
Helena Bylin
2   Department of Molecular and Clinical Medicine, Division of Occupational and Environmental Medicine, Linköping, Sweden
,
Per Leanderson
2   Department of Molecular and Clinical Medicine, Division of Occupational and Environmental Medicine, Linköping, Sweden
,
Arina Richter
3   Department of Medicine and Care, Division of Cardiology
,
Magnus Grenegård
1   Department of Medicine and Care, Division of Pharmacology
,
Torbjörn Bengtsson
1   Department of Medicine and Care, Division of Pharmacology
› InstitutsangabenGrant support: This study was supported by the Swedish Research Council (grant number 12668), Trygg Hansa Research Foundation, The Strategic Research Programmes “Inflammation” and “Cardiovascular Inflammation Research Center” of Linköping University, Heart Foundation of Linköping University, Östergötland Country Council Committee for Medical Research and Development, Medical Research Council of Southeast Sweden.
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Publikationsverlauf

Received: 10. Juni 2004

Accepted after major revision: 13. April 2005

Publikationsdatum:
05. Dezember 2017 (online)

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Summary

There is increasing evidence that Chlamydia pneumoniae is linked to atherosclerosis and thrombosis. In this regard, we have recently shown that C. pneumoniae stimulates platelet aggregation and secretion, which may play an important role in the progress of atherosclerosis and in thrombotic vascular occlusion. The aims of the present study were to investigate the effects of C. pneumoniae on platelet-mediated formation of reactive oxygen species (ROS) and oxidation of low-density lipoprotein (LDL) in vitro. ROS production was registered as changes in 2‘,7’-dichlorofluorescin- fluorescence in platelets with flow cytometry. LDL-oxidation was determined by measuring thiobarbituric acid reactive substances (TBARs). We found that C. pneumoniae stimulated platelet production of ROS. Polymyxin B treatment of C. pneumoniae, but not elevated temperature, abolished the stimulatory effects on platelet ROS- production, which suggests that chlamydial lipopolysaccharide has an important role. Inhibition of nitric oxide synthase with nitro-L-arginine, lipoxygenase with 5,8,11-eicosatriynoic acid and protein kinase C with GF 109203X significantly lowered the production of radicals. In contrast, inhibition of NADPH-oxidase with di-phenyleneiodonium (DPI) did not affect the C. pneumoniae induced ROS-production. These findings suggest that the activities of nitric oxide synthase and lipoxygenase are the sources for ROS and that the generation is dependent of the activity of protein kinase C. The C. pneumoniae-induced ROS-production in platelets was associated with an extensive oxidation of LDL, which was significantly higher compared to the effect obtained by separate exposure of LDL to C. pneumoniae or platelets. In conclusion, C. pneumoniae interaction with platelets leading to aggregation, ROS-production and oxidative damage on LDL, may play a crucial role in the development of atherosclerotic cardiovascular disease.

Keywords

Atherosclerosis - bacteria-cell interaction - oxygen radical - LPS - thrombosis
 

  • References

  • 1 Mygind T, Birkelund S, Falk E. et al. Evaluation of real-time quantitative PCR for identification and quantification of Chlamydia pneumoniae by comparison with immunohistochemistry. J Microbiol Methods 2001; 46: 241-51.
    CrossrefPubMedGoogle Scholar
  • 2 Saikku P. Chlamydia pneumoniae and atherosclerosis– an update. Scand J Infect Dis Suppl 1997; 104: 53-6.
    PubMedGoogle Scholar
  • 3 Larsen MM, Moern B, Fuller A. et al. Chlamydia pneumoniae and cardiovascular disease. Med J Aust 2002; 177: 558-62.
    CrossrefPubMedGoogle Scholar
  • 4 Leinonen M, Saikku P. Evidence for infectious agents in cardiovascular disease and atherosclerosis. Lancet Infect Dis 2002; 2: 11-7.
    CrossrefPubMedGoogle Scholar
  • 5 Maass M, Jahn J, Gieffers J. et al. Detection of Chlamydia pneumoniae within peripheral blood monocytes of patients with unstable angina or myocardial infarction. J Infect Dis 2000; 181 (Suppl. 03) S449-51.
    CrossrefPubMedGoogle Scholar
  • 6 Gaydos CA. Growth in vascular cells and cytokine production by Chlamydia pneumoniae . J Infect Dis 2000; 181 (Suppl. 03) S473-8.
    CrossrefPubMedGoogle Scholar
  • 7 Kalayoglu MV, Indrawati Morrison RP. et al. Chlamydial virulence determinants in atherogenesis: the role of chlamydial lipopolysaccharide and heat shock protein 60 in macrophage-lipoprotein interactions. J Infect Dis 2000; 181 (Suppl. 03) S483-9.
    CrossrefPubMedGoogle Scholar
  • 8 Kalayoglu MV, Hoerneman B, LaVerda D. et al. Cellular oxidation of low-density lipoprotein by Chlamydia pneumoniae . J Infect Dis 1999; 180: 780-90.
    CrossrefPubMedGoogle Scholar
  • 9 Kalayoglu MV, Byrne GI. Induction of macrophage foam cell formation by Chlamydia pneumoniae . J Infect Dis 1998; 177: 725-9.
    CrossrefPubMedGoogle Scholar
  • 10 Molestina RE, Miller RD, Ramirez JA. et al. Infection of human endothelial cells with Chlamydia pneumoniae stimulates transendothelial migration of neutrophils and monocytes. Infect Immun 1999; 67: 1323-30.
    CrossrefPubMedGoogle Scholar
  • 11 Hirono S, Dibrov E, Hurtado C. et al. Chlamydia pneumoniae stimulates proliferation of vascular smooth muscle cells through induction of endogenous heat shock protein 60. Circ Res 2003; 93: 710-6.
    CrossrefPubMedGoogle Scholar
  • 12 Kalvegren H, Majeed M, Bengtsson T. Chlamydia pneumoniae binds to platelets and triggers P-selectin expression and aggregation: A causal role in cardiovascular disease?. Arterioscler Thromb Vasc Biol 2003; 23: 1677-83.
    CrossrefPubMedGoogle Scholar
  • 13 Zielinski T, Wachowicz B, Saluk-Juszczak J. et al. The generation of superoxide anion in blood platelets in response to different forms of Proteus mirabilis lipopolysaccharide: effects of staurosporin, wortmannin, and indomethacin. Thromb Res 2001; 103: 149-55.
    CrossrefPubMedGoogle Scholar
  • 14 Marcus AJ, Silk ST, Safier LB. et al. Superoxide production and reducing activity in human platelets. J Clin Invest 1977; 59: 149-58.
    CrossrefPubMedGoogle Scholar
  • 15 Krotz F, Sohn HY, Pohl U. Reactive oxygen species. players in the platelet game. Arterioscler Thromb Vasc Biol 2004; 24: 1988-96.
    CrossrefPubMedGoogle Scholar
  • 16 Harrison D, Griendling KK, Landmesser U. et al. Role of oxidative stress in atherosclerosis. Am J Cardiol 2003; 91: 7A-11A.
    CrossrefPubMedGoogle Scholar
  • 17 Taniyama Y, Griendling KK. Reactive oxygen species in the vasculature. molecular and cellular mechanisms. Hypertension 2003; 42: 1075-81.
    CrossrefPubMedGoogle Scholar
  • 18 Maytin M, Leopold J, Loscalzo J. Oxidant stress in the vasculature. Curr Atheroscler Rep 1999; 1: 156-64.
    CrossrefPubMedGoogle Scholar
  • 19 Chamulitrat W, Mason RP. Lipid peroxyl radical intermediates in the peroxidation of polyunsaturated fatty acids by lipoxygenase. Direct electron spin resonance investigations. J Biol Chem 1989; 264: 20968-73.
    CrossrefPubMedGoogle Scholar
  • 20 Jahn B, Hansch GM. Oxygen radical generation in human platelets: dependence on 12-lipoxygenase activity and on the glutathione cycle. Int Arch Allergy Appl Immunol 1990; 93: 73-9.
    CrossrefPubMedGoogle Scholar
  • 21 Seno T, Inoue N, Gao D. et al. Involvement of NADH/NADPH oxidase in human platelet ROS production. Thromb Res 2001; 103: 399-409.
    CrossrefPubMedGoogle Scholar
  • 22 Iuliano L, Colavita AR, Leo R. et al. Oxygen free radicals and platelet activation. Free Radic Biol Med 1997; 22: 999-1006.
    CrossrefPubMedGoogle Scholar
  • 23 Muruganandam A, Mutus B. Isolation of nitric oxide synthase from human platelets. Biochim Biophys Acta 1994; 1200: 1-6.
    CrossrefPubMedGoogle Scholar
  • 24 Caccese D, Pratico D, Ghiselli A. et al. Superoxide anion and hydroxyl radical release by collagen-induced platelet aggregation–role of arachidonic acid metabolism. Thromb Haemost 2000; 83: 485-90.
    Artikel in Thieme ConnectPubMedGoogle Scholar
  • 25 Redecke V, Dalhoff K, Bohnet S. et al. Interaction of Chlamydia pneumoniae and human alveolar macrophages: infection and inflammatory response. Am J Respir Cell Mol Biol 1998; 19: 721-7.
    CrossrefPubMedGoogle Scholar
  • 26 van Kuppeveld FJ, van der, Logt JT, Angulo AF. et al. Genus- and species-specific identification of mycoplasmas by 16S rRNA amplification. Appl Environ Microbiol 1992; 58: 2606-15.
    CrossrefPubMedGoogle Scholar
  • 27 Bengtsson T, Zalavary S, Stendahl O. et al. Release of oxygen metabolites from chemoattractant-stimulated neutrophils is inhibited by resting platelets: role of extracellular adenosine and actin polymerization. Blood 1996; 87: 4411-23.
    CrossrefPubMedGoogle Scholar
  • 28 Myhre O, Andersen JM, Aarnes H. et al. Evaluation of the probes 2’,7’-dichlorofluorescin diacetate, luminol, and lucigenin as indicators of reactive species formation. Biochem Pharmacol 2003; 65: 1575-82.
    CrossrefPubMedGoogle Scholar
  • 29 da Silva EL, Tsushida T, Terao J. Inhibition of mammalian 15-lipoxygenase-dependent lipid peroxidation in low-density lipoprotein by quercetin and quercetin monoglucosides. Arch Biochem Biophys 1998; 349: 313-20.
    CrossrefPubMedGoogle Scholar
  • 30 Folcik VA, Aamir R, Cathcart MK. Cytokine modulation of LDL oxidation by activated human monocytes. Arterioscler Thromb Vasc Biol 1997; 17: 1954-61.
    CrossrefPubMedGoogle Scholar
  • 31 Klinger MH. Platelets and inflammation. Anat Embryol (Berl) 1997; 196: 1-11.
    CrossrefPubMedGoogle Scholar
  • 32 Benov LT, Fridovich I. Escherichia coli expresses a copper- and zinc-containing superoxide dismutase. J Biol Chem 1994; 269: 25310-4.
    CrossrefPubMedGoogle Scholar
  • 33 Karavolos MH, Horsburgh MJ, Ingham E. et al. Role and regulation of the superoxide dismutases of Staphylococcus aureus. Microbiology 2003; 149: 2749-58.
    CrossrefPubMedGoogle Scholar
  • 34 Pulcinelli FM, Pignatelli P, Violi F. et al. Platelets and oxygen radicals: mechanisms of functional modulation. Haematologica 2001; 86 (Suppl. 112) 31-4.
    PubMedGoogle Scholar
  • 35 Pignatelli P, Pulcinelli FM, Lenti L. et al. Hydrogen peroxide is involved in collagen-induced platelet activation. Blood 1998; 91: 484-90.
    CrossrefPubMedGoogle Scholar
  • 36 Lufrano M, Balazy M. Interactions of peroxynitrite and other nitrating substances with human platelets: the role of glutathione and peroxynitrite permeability. Biochem Pharmacol 2003; 65: 515-23.
    CrossrefPubMedGoogle Scholar
  • 37 Pou S, Pou WS, Bredt DS. et al. Generation of superoxide by purified brain nitric oxide synthase. J Biol Chem 1992; 267: 24173-6.
    CrossrefPubMedGoogle Scholar
  • 38 Chakrabarti S, Clutton P, Varghese S. et al. Glycoprotein IIb/IIIa inhibition enhances platelet nitric oxide release. Thromb Res 2004; 113: 225-33.
    CrossrefPubMedGoogle Scholar
  • 39 Denicola A, Souza JM, Radi R. Diffusion of peroxynitrite across erythrocyte membranes. Proc Natl Acad Sci U S A 1998; 95: 3566-71.
    CrossrefPubMedGoogle Scholar
  • 40 Khairutdinov RF, Coddington JW, Hurst JK. Permeation of phospholipid membranes by peroxynitrite. Biochemistry 2000; 39: 14238-49.
    CrossrefPubMedGoogle Scholar
  • 41 Romero N, Denicola A, Souza JM. et al. Diffusion of peroxynitrite in the presence of carbon dioxide. Arch Biochem Biophys 1999; 368: 23-30.
    CrossrefPubMedGoogle Scholar
  • 42 Sucu N, Unlu A, Tamer L. et al. 3-Nitrotyrosine in atherosclerotic blood vessels. Clin Chem Lab Med 2003; 41: 23-5.
    CrossrefPubMedGoogle Scholar
  • 43 Yamaguchi Y, Matsuno S, Kagota S. et al. Peroxynitrite- mediated oxidative modification of low-density lipoprotein by aqueous extracts of cigarette smoke and the preventive effect of fluvastatin. Atherosclerosis 2004; 172: 259-65.
    CrossrefPubMedGoogle Scholar
  • 44 Li J, Li W, Su J. et al. Peroxynitrite induces apoptosis in rat aortic smooth muscle cells: possible relation to vascular diseases. Exp Biol Med 2004; 229: 264-9.
    CrossrefPubMedGoogle Scholar
  • 45 Rund S, Lindner B, Brade H. et al. Structural analysis of the lipopolysaccharide from Chlamydia trachomatis serotype L2. J Biol Chem 1999; 274: 16819-24.
    CrossrefPubMedGoogle Scholar
  • 46 Ogawa T. Chemical structure of lipid A from Porphyromonas (Bacteroides) gingivalis lipopolysaccharide. FEBS Lett 1993; 332: 197-201.
    CrossrefPubMedGoogle Scholar
  • 47 Endo Y, Shibazaki M, Nakamura M. et al. Contrasting effects of lipopolysaccharides (endotoxins) from oral black-pigmented bacteria and Enterobacteriaceae on platelets, a major journal-title of serotonin, and on histamine- forming enzyme in mice. J Infect Dis 1997; 175: 1404-12.
    CrossrefPubMedGoogle Scholar
  • 48 Grabarek J, Timmons S, Hawiger J. Modulation of human platelet protein kinase C by endotoxic lipid A. J Clin Invest 1988; 82: 964-71.
    CrossrefPubMedGoogle Scholar
  • 49 Berliner JA, Heinecke JW. The role of oxidized lipoproteins in atherogenesis. Free Radic Biol Med 1996; 20: 707-27.
    CrossrefPubMedGoogle Scholar
  • 50 Memon RA, Staprans I, Noor M. et al. Infection and inflammation induce LDL oxidation in vivo. Arterioscler Thromb Vasc Biol 2000; 20: 1536-42.
    CrossrefPubMedGoogle Scholar