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Oxidative stress induces myeloperoxidase expression in endocardial endothelial cells from patients with chronic heart failure

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Abstract

Increased oxidative stress has been implicated in the pathogenesis of a number of cardiovascular diseases. Recent findings suggest that myeloperoxidase (MPO) may play a key role in the initiation and maintenance of chronic heart failure (CHF) by contributing to the depletion of the intracellular reservoir of nitric oxide (NO). NO consumption through MPO activity may lead to protein chlorination or nitration, leading to tissue damage. Primary cultures of human endocardial endothelial cells (EEC) obtained at heart transplantation of patients with CHF and human umbilical vein endothelial cells (HUVEC) were subjected to oxidative stress by incubation with hydrogen peroxide at non lethal (60 µM) dose for different exposure times (3 and 6 h). Treated and control cells were tested by immunohistochemistry and RT-PCR for MPO and 3-chlorotyrosine expression. Both endothelial cell types expressed myeloperoxidase following oxidative stress, with higher levels in EEC. Moreover, 3-chlorotyrosine accumulation in treated cells alone indicated the presence of MPO-derived hypochlorous acid. Immunohistochemistry on sections from post-infarcted heart confirmed in vivo the endothelial positivity to MPO, 3-chlorotyrosine and, to a minor extent, nitrotyrosine. Immunohistochemical observations were confirmed by detection of MPO mRNA in both stimulated EEC and HUVEC cells. This study demonstrates for the first time that EEC can express MPO after oxidative stress, both in vitro and in vivo, followed by accumulation of 3-chlorotyrosine, an end product of oxidative stress. Deregulation of endothelial functions may contribute to the development of a number of cardiovascular diseases, including CHF. The results also highlight the notion that endothelium is not only a target but also a key player in oxidative-driven cardiovascular stress.

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References

  1. Baldus S, Eiserich JP, Mani A, Castro L, Figueroa M, Chumley P, Ma W, Tousson A, White CR, Bullard DC, Brennan ML, Lusis AJ, Moore KP, Freeman BA (2001) Endothelial transcytosis of myeloperoxidase confers specificity to vascular ECM proteins as targets of tyrosine nitration. J Clin Invest 108:1759–1770

    PubMed  CAS  Google Scholar 

  2. Baldus S, Heitzer T, Eiserich JP, Lau D, Mollnau H, Ortak M, Petri S, Goldmann B, Duchstein HJ, Berger J, Helmchen U, Freeman BA, Meinertz T, Munzel T (2004) Myeloproxidase enhances nitric oxide catabolism during myocardial ischemia and reperfusion. Free Radic Biol Med 37:902–911

    Article  PubMed  CAS  Google Scholar 

  3. Baldus S, Rudolph V, Roiss M, Ito WD, Rudolph TK, Eiserich JP, Sydow K, Lau D, Szoks K, Klinke A, Kubala L, Berglund L, Schrepfer S, Deuse T, Haddad M, Risius T, Klemm H, Reichenspurner HC, Meinertz T, Heitzer T (2006) Heparins increase endothelial nitric oxide bioavailability by liberating vessel-immobilized myeloperoxidase. Circulation 113:1871–1878

    Article  PubMed  CAS  Google Scholar 

  4. Berry CE, Hare JM (2004) Xanthine oxidoreductase in the cardiovascular system: molecular mechanisms and pathophysiologic implications. J Physiol 555:589–606

    Article  PubMed  CAS  Google Scholar 

  5. Bradley JR, Johnson DR, Pober JS (1993) Endothelial activation by hydrogen peroxide. Am J Pathol 142:1598–1609

    PubMed  CAS  Google Scholar 

  6. Cai H (2005) Hydrogen peroxide regulation of endothelial function: Origin, mechanism and consequences. Cardiovasc Res 68:26–36

    Article  PubMed  CAS  Google Scholar 

  7. Cappello F, Di Stefano A, David S, Rappa F, Anzalone R, La Rocca G, D’Anna SE, Magno F, Donner CF, Balbi B, Zummo G (2006) Hsp60 and Hsp10 down-regulation predicts bronchial epithelial carcinogenesis in smokers with chronic obstructive pulmonary disease. Cancer 107:2417–2424

    Article  PubMed  CAS  Google Scholar 

  8. Chen X, Niroomand F, Liu Z, Zankl A, Katus HA, Jahn L, Tiefenbacher CP (2006) Expression of nitric oxide related enzymes in coronary heart disease. Basic Res Cardiol 101:346–353

    Article  PubMed  CAS  Google Scholar 

  9. Clancy RM, Leszczynskapiziak J, Abramson SB (1992) Nitric-oxide, an endothelial-cell relaxation factor, inhibits neutrophil superoxide anion production via a direct action on the NADPH oxidase. J Clin Invest 90:1116–1121

    Article  PubMed  CAS  Google Scholar 

  10. Cote CG, Yu FS, Zulueta JJ, Vosatka RJ, Hassoun PM (1996) Regulation of intracellular xanthine oxidase by endothelial-derived nitric oxide. Am J Physiol Lung Cell Mol Physiol 15:L869–L874

    Google Scholar 

  11. Daugherty A, Dunn JL, Rateri DL, Heinecke JW (1994) Myeloperoxidase, a catalyst for lipoprotein oxidation, is expressed in human atherosclerotic lesions. J Clin Invest 94:437–444

    Article  PubMed  CAS  Google Scholar 

  12. Drummond GR, Cai H, Davis ME, Ramasamy S, Harrison DG (2000) Transcriptional and posttranscriptional regulation of endothelial nitric oxide synthase expression by hydrogen peroxide. Circ Res 86:347–354

    PubMed  CAS  Google Scholar 

  13. Eu JP, Hare JM, Hess DT, Skaf M, Sun J, Cordenas-Navina I, Sun QA, Dewhirst M, Meissner G, Stamler JS (2003) Concerted regulation of skeletal muscle contractility by oxygen tension and endogenous nitric oxide. Proc Natl Acad Sci USA 100:15229–15234

    Article  PubMed  CAS  Google Scholar 

  14. Frantz S, Brandes RP, Hu K, Rammelt K, Wolf J, Scheuermann H, Ertl G, Bauersachs J (2006) Left ventricular remodeling after myocardial infarction in mice with targeted deletion of the NADPH oxidase subunit gp91PHOX. Basic Res Cardiol 101:127–132

    Article  PubMed  CAS  Google Scholar 

  15. Garcia SC, Pomblum V, Gams E, Langenbach MR, Schipke JD (2007) Independency of myocardial stunning of endothelial stunning? Basic Res Cardiol 102:359–367

    Article  PubMed  CAS  Google Scholar 

  16. Gaut JP, Yeh GC, Tran HD, Byun J, Henderson JP, Richter GM, Brennan ML, Lusis AJ, Belaaouaj A, Hotchkiss RS, Heinecke JW (2001) Neutrophils employ the myeloperoxidase system to generate antimicrobial brominating and chlorinating oxidants during sepsis. Proc Natl Acad Sci USA 98:11961–11966

    Article  PubMed  CAS  Google Scholar 

  17. Giordano FJ (2005) Oxygen, oxidative stress, hypoxia, and heart failure. J Clin Invest 115:500–508

    PubMed  CAS  Google Scholar 

  18. Green PS, Mendez AJ, Jacob JS, Crowley JR, Growdon W, Hyman BT, Heinecke JW (2004) Neuronal expression of myeloperoxidase is increased in Alzheimer’s disease. J Neurochem 90:724–733

    Article  PubMed  CAS  Google Scholar 

  19. Hare JM, Stamler JS (2005) NO/redox disequilibrium in the failing heart and cardiovascular system J Clin Invest 115:509–517

    PubMed  CAS  Google Scholar 

  20. Hazen SL, Heinecke JW (1997) 3-Chlotyrosine, a specific marker of myeloperoxidase-catalyzed oxidation, is markedly elevated in low density lipoprotein isolated from human atherosclerotic intima. J Clin Invest 99:2075–2081

    Article  PubMed  CAS  Google Scholar 

  21. Jaffe EA, Nachman RL, Becker CG, Minick CR (1973) Culture of human endothelial cells derived from umbilical veins: identification by morphologic and immunologic criteria. J Clin Invest 52:2745–2756

    Article  PubMed  CAS  Google Scholar 

  22. Khan SA, Lee K, Minhas KM, Gonzalez DR, Raju SD, Tejani AD, Li D, Berkowitz DE, Hare JM (2004) Neuronal nitric oxide synthase negatively regulates xanthine oxidoreductase inhibition of cardiac excitation-contraction coupling. Proc Natl Acad Sci USA 101:15944–15948

    Article  PubMed  CAS  Google Scholar 

  23. Klebanoff SJ (2005) Myeloperoxidase: friend and foe. J Leukoc Biol 77:598–625

    Article  PubMed  CAS  Google Scholar 

  24. Kumar AP, Piedrafita FJ, Reynolds WF (2004) Peroxisome proliferator-activated receptor γ ligands regulate myeloperoxidase expression in macrophages by an estrogen-dependent mechanism involving the –463GA promoter polymorphism. J Biol Chem 279:8300–8315

    Article  PubMed  CAS  Google Scholar 

  25. La Rocca G, Anzalone R, Magno F, Farina F, Cappello F, Zummo G (2007) Cigarette smoke exposure inhibits extracellular MMP-2 (gelatinase A) activity in human lung fibroblasts. Resp Res 8:23

    Article  Google Scholar 

  26. Landmesser U, Dikalov S, Price SR, McCann L, Fukai T, Holland SM, Mitch WE, Harrison DG (2003) Oxidation of tetrahydrobiopterin leads to uncoupling of endothelial cell nitric oxide synthase in hypertension. J Clin Invest 111:1201–1209

    PubMed  CAS  Google Scholar 

  27. Lau D, Baldus S (2006) Myeloperoxidase and its contributory role in inflammatory vascular disease. Pharmacol Ther 111:16–26

    Article  PubMed  CAS  Google Scholar 

  28. Linke A, Recchia F, Zhang X, Hintze TH (2003) Acute and chronic endothelial dysfunction: implications for the development of heart failure. Heart Fail Rev 8:87–97

    Article  PubMed  CAS  Google Scholar 

  29. Liu X, Zweier JL (2001) A real-time electrochemical technique for measurement of cellular hydrogen peroxide generation and consumption: evaluation in human polymorphonuclear leukocytes. Free Radic Biol Med 31:894–901

    Article  PubMed  CAS  Google Scholar 

  30. Maciag T, Hoover GA, Stemerman MB, Weinstein R (1981) Serial propagation of human endothelial cells in vitro. J Cell Biol 91:420–426

    Article  PubMed  CAS  Google Scholar 

  31. Marin V, Kaplanski G, Gres S, Farnarier C, Bongrand P (2001) Endothelial cell culture: protocol to obtain and cultivate human umbilical endothelial cells. J Immunol Methods 254:183–190

    Article  PubMed  CAS  Google Scholar 

  32. Moens AL, Kass DA (2006) Tetrahydrobiopterin and cardiovascular disease. Arterioscler Thromb Vasc Biol 26:2439–2444

    Article  PubMed  CAS  Google Scholar 

  33. Nicholls SJ, Hazen SL (2005) Myeloperoxidase and cardiovascular disease. Arterioscler Thromb Vasc Biol 25:1102–1111

    Article  PubMed  CAS  Google Scholar 

  34. Nie H, Wu JL, Zhang M, Xu J, Zou MH (2006) Endothelial nitric oxide synthase-dependent tyrosine nitration of prostacyclin synthase in diabetes in vivo. Diabetes 55:3133–3141

    Article  PubMed  CAS  Google Scholar 

  35. Podrez EA, Abu-Soud HM, Hazen SL (2000) Myeloperoxidase-generated oxidants and atherosclerosis. Free Radic Biol Med 28:1717–1725

    Article  PubMed  CAS  Google Scholar 

  36. Ricciardolo F, Caramori G, Ito K, Capelli A, Brun P, Abatangelo G, Papi A, Chung KF, Adcock I, Barnes PJ, Donner CF, Rossi A, Di Stefano A (2005) Nitrosative stress in the bronchial mucosa of severe chronic obstructive pulmonary disease. J Allergy Clin Immunol 115:779–785

    Article  PubMed  Google Scholar 

  37. Roy S, Khanna S, Sehn CK (2008) Redox regulation of the VEGF signaling path and tissue vascularization: Hydrogen peroxide, the common link between physical exercise and cutaneous wound healing. Free Rad Biol Med 44:180–192

    Article  PubMed  CAS  Google Scholar 

  38. Salavej P, Spalteholz H, Arnhold J (2006) Modification of amino acid residues in human serum albumin by myeloperoxidase. Free Radic Biol Med 40:516–525

    Article  PubMed  CAS  Google Scholar 

  39. Shinyashiki M, Pan CJG, Lopez BE, Fukuto JM (2004) Inhibition of the yeast metal reductase heme protein Fre1 by nitric oxide (NO): A model for inhibition of NADPH oxidase by NO. Free Radic Biol Med 37:713–723

    Article  PubMed  CAS  Google Scholar 

  40. Stilli D, Lagrasta C, Berni R, Bocchi L, Savi M, Delucchi F, Graiani G, Monica M, Maestri R, Baruffi S, Rossi S, Macchi E, Musso E, Quaini F (2007) Preservation of ventricular performance at early stages of diabetic cardiomyopathy involves changes in myocyte size, number and intercellular coupling. Basic Res Cardiol 102:488–499

    Article  PubMed  CAS  Google Scholar 

  41. Sugiyama S, Okada Y, Sukhova GK, Birmani R, Heinecke JW, Libby P (2001) Macrophage myeloperoxidase regulation by granulocyte macrophage colony-simulating factor in human atherosclerosis and implications in acute coronary syndromes. Am J Pathol 158:879–891

    PubMed  CAS  Google Scholar 

  42. Suhara T, Fukuo K, Sugimoto T, Morimoto S, Nakahashi T, Hata S, Shimizu M, Ogihara T (1998) Hydrogen peroxide induces up-regulation of Fas in human endothelial cells. J Immunol 160:4042–4047

    PubMed  CAS  Google Scholar 

  43. Tang WHW, Brennan M-L, Philip K, Tong W, Mann S, Van Lente F, Hazen SL (2006) Plasma myeloperoxidase levels in patients with chronic heart failure. Am J Cardiol 98:796–799

    Article  PubMed  CAS  Google Scholar 

  44. Thomas SR, Chen K, Keaney, JF Jr (2002) Hydrogen peroxide activates endothelial nitric-oxide synthase through coordinated phosphorylation and dephosphorylation via a phosphoinositide 3-kinase-dependent signaling pathway. J Biol Chem 277:6017–6024

    Article  PubMed  CAS  Google Scholar 

  45. Tiruppathi C, Naqvi T, Wu Y, Vogel SM, Minshall RD, Malik AB (2004) Albumin mediates the transcytosis of myeloperoxidase by means of caveolae in endothelial cells. Proc Natl Acad Sci USA 101:7699–7704

    Article  PubMed  CAS  Google Scholar 

  46. Van der Vliet A, Eiserich JP, Shigenaga MK, Cross CE (1999) Reactive nitrogen species and tyrosine nitration in the respiratory tract. epiphenomena or a pathobiologic mechanism of disease? Am J Respir Crit Care Med 160:1–9

    PubMed  Google Scholar 

  47. Vita JA, Brennan ML, Goke N, Mann SA, Goormastic M, Shishehbor MH, Penn MS, Keaney JF Jr, Hazen SL (2004) Serum myeloperoxidase levels independently predict endothelial dysfunction in humans. Circulation 110:1134–1139

    Article  PubMed  CAS  Google Scholar 

  48. Winterbourn CC, Kettle AJ (2000) Biomarkers of myeloperoxidase-derived hypochlorous acid. Free Radic Biol Med 5:403–409

    Article  Google Scholar 

  49. Zhen J, Lu H, Wang XQ, Vaziri ND, Zhou XJ (2008) Upregulation of endothelial and inducible nitric oxide synthase expression by reactive oxygen species. Am J Hypertens 21:28–34

    Article  PubMed  CAS  Google Scholar 

  50. Zou MH (2007) Peroxynitrite and protein tyrosine nitration of prostacyclin synthase. Prostaglandins Other Lipid Mediat 82:119–127

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

We thank Rosemary Allpress for her revision of the English language. We thank Antonella Chiara, Santina Di Gangi and Giusy Scaduto for the valuable support during the experimental work. This work was supported by Fondazione S. Maugeri, IRCCS, Ricerca corrente, and Italian Ministry of University (Ex 60% to GZ, FC, FF) grants. The authors declare no conflict of interests.

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Authors and Affiliations

  1. Sezione di Anatomia Umana, Dipto. di Medicina Sperimentale, Università degli Studi di Palermo, Via del Vespro 129, 90127, Palermo, Italy

    Giampiero La Rocca, Rita Anzalone, Francesca Magno, Simona Corrao, Tiziana Loria, Felicia Farina, Francesco Cappello & Giovanni Zummo

  2. Laboratorio di Citoimmunopatologia Apparato Cardio-Respiratorio, Fondazione S. Maugeri, IRCCS, Veruno, NO, Italy

    Antonino Di Stefano & Marilena Colombo

  3. Divisione di Cardiologia, Fondazione S. Maugeri, IRCCS, Veruno, NO, Italy

    Ermanno Eleuteri & Pantaleo Giannuzzi

  4. Dipto. di Patologia Umana, Università degli Studi di Palermo, Palermo, Italy

    Anna Martorana

  5. Istituto di Ostetricia e Ginecologia, Università degli Studi di Palermo, Palermo, Italy

    Claudio Di Gangi

  6. Divisione di Cardiochirurgia, Ospedale S. Giovanni Battista, Turin, Italy

    Fabrizio Sansone, Francesco Patanè & Mauro Rinaldi

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Correspondence to Giampiero La Rocca.

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G. La Rocca and A. Di Stefano contributed equally to the current work.

Returned for 1. Revision: 7 January 2008 1. Revision received: 18 June 2008

Returned for 2. Revision: 17 July 2008 2. Revision received: 17 October 2008

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La Rocca, G., Di Stefano, A., Eleuteri, E. et al. Oxidative stress induces myeloperoxidase expression in endocardial endothelial cells from patients with chronic heart failure. Basic Res Cardiol 104, 307–320 (2009). https://doi.org/10.1007/s00395-008-0761-9

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