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Amino Acids

, Volume 40, Issue 4, pp 1127–1137 | Cite as

A pro-survival effect of polyamine depletion on norepinephrine-mediated apoptosis in cardiac cells: role of signaling enzymes

  • Silvia Cetrullo
  • Benedetta TantiniEmail author
  • Annalisa Facchini
  • Carla Pignatti
  • Claudio Stefanelli
  • Claudio Marcello Caldarera
  • Flavio Flamigni
Original Article

Abstract

Recent studies report that the primary transmitter of sympathetic nervous system norepinephrine (NE), which is actively produced in failing human heart, is able to induce apoptosis of rat cardiomyocytes. Apoptotic cell death of cardiomyocytes is involved in several cardiovascular diseases including ischemia, hypertrophy and heart failure, therefore representing a potential therapeutic target. The natural occurring polyamines, putrescine, spermidine and spermine, are biogenic amines involved in many cellular processes, including apoptosis. Thus, we have studied the involvement of polyamines in the apoptosis of cardiac cells induced by the treatment with NE. The results indicate that NE caused an early induction of the activity of ornithine decarboxylase (ODC), the first enzyme in polyamine biosynthesis, followed by a later increase of apoptotic cell death. This effect was prevented in the presence of α-difluoromethylornithine, an irreversible inhibitor of ODC. Moreover, the study of some key signal transduction pathways revealed an involvement of AMP-activated protein kinase, AKT and p38 mitogen-activated protein kinases, in the modulation by polyamines of the response of cardiomyocytes to NE. In fact, polyamine-depleted cells showed an altered activation pattern of these kinases that may contrast apoptosis and appeared to result from a differential effect on the specific phosphatases that dephosphorylate and switch off these signaling proteins. In conclusion, these results indicate that in cardiac cells polyamines are involved in the execution of the death program activated by NE, and suggest that their apoptosis facilitating action is mediated by a network of specific phosphatases and kinases.

Keywords

Norepinephrine Cardiac cells Apoptosis Polyamines Kinases Phosphatases 

Notes

Acknowledgments

The excellent technical assistance of Maddalena Zini is acknowledged. This work was supported by grants from University of Bologna (R.F.O.), Istituto Nazionale per le Ricerche Cardiovascolari (INRC) and Compagnia di San Paolo, Torino, Italy.

References

  1. Abbate A, Scarpa S, Santini D, Palleiro J, Vasaturo F, Miller J, Morales C, Vetrovec GW, Baldi A (2006) Myocardial expression of survivin, an apoptosis inhibitor, in aging and heart failure: an experimental study in the spontaneously hypertensive rat. Int J Cardiol 11(3):371–376CrossRefGoogle Scholar
  2. Averill-Bates D, Ke Q, Tanel A, Roy J, Fortier G, Agostinelli E (2008) Mechanism of cell death induced by spermine and amine oxidase in mouse melanoma cells. Int J Oncol 32:79–88PubMedGoogle Scholar
  3. Bhattacharya S, Ray RM, Viar MJ, Johnson RL (2003) Polyamines are required for activation of c-Jun NH2-terminal kinase and apoptosis in response to TNF-alpha in IEC-6 cells. Am J Physiol Gastrointest Liver Physiol 285:G980–G991PubMedGoogle Scholar
  4. Bhattacharya S, Ray RM, Johnson LR (2005) Decreased apoptosis in polyamine depleted IEC-6 cells depends on Aktmediated NF-kappaB activation but not GSK3beta activity. Apoptosis 10:759–776PubMedCrossRefGoogle Scholar
  5. Buerke M, Murohara T, Skurk C, Nuss C, Tomaselli K, Lefer AM (1995) Cardioprotective effect of insulin-like growth factor I in myocardial ischemia followed by reperfusion. Proc Natl Acad Sci USA 92:8031–8035PubMedCrossRefGoogle Scholar
  6. Capano M, Crompton M (2006) Bax translocates to mitochondria of heart cells during simulated ischaemia: involvement of AMP-activated and p38 mitogen-activated protein kinases. Biochem J 1:57–64Google Scholar
  7. Cetrullo S, Facchini A, Stanic I, Tantini B, Pignatti C, Caldarera CM et al (2010) Difluoromethylornithine inhibits hypertrophic, pro-fibrotic and pro-apoptotic actions of aldosterone in cardiac cells. Amino Acids 38:525–531PubMedCrossRefGoogle Scholar
  8. Chun TY, Pratt JH (2005) Aldosterone increases plasminogen activator inhibitor-1 synthesis in rat cardiomyocytes. Mol Cell Endocrinol 239:55–61PubMedCrossRefGoogle Scholar
  9. Clerk A, Cullingford TE, Fuller SJ, Giraldo A, Markou T, Pikkarainen S et al (2007) Signaling pathways mediating cardiac myocyte gene expression in physiological and stress responses. J Cell Physiol 212:311–322PubMedCrossRefGoogle Scholar
  10. Colucci WS, Sawyer DB, Singh K, Communal C (2000) Adrenergic overload and apoptosis in heart failure: implications for therapy. J Card Fail 6:1–7PubMedCrossRefGoogle Scholar
  11. Communal C, Singh K, Pimentel DR, Colucci WS (1998) Norepinephrine stimulates apoptosis in adult rat ventricular myocytes by activation of the beta-adrenergic pathway. Circulation 98:1329–1334PubMedGoogle Scholar
  12. Cornwell T, Mehta P, Shenolikar S (1986) Polyamine stimulation of protein phosphatase-2A from rat liver using a non-protein phosphoester substrate. J Cyclic Nucleotide Protein Phosphor Res 11:373–382PubMedGoogle Scholar
  13. Czerski L, Nuñez G (2004) Apoptosome formation and caspase activation: is it different in the heart? J Mol Cell Cardiol 37:643–652PubMedCrossRefGoogle Scholar
  14. Davies SP, Helps NR, Cohen PT, Hardie DG (1995) 5-AMP inhibits dephosphorylation, as well as promoting phosphorylation, of the AMP-activated protein kinase. Studies using bacterially expressed human protein phosphatase-2C alpha and native bovine protein phosphatase-2AC. FEBS Lett 377:421–425PubMedCrossRefGoogle Scholar
  15. Dyck JRB, Lopaschuk GD (2006) AMPK alterations in cardiac physiology and pathology: enemy or ally? J Physiol 574:95–112PubMedCrossRefGoogle Scholar
  16. Fan W J, van Vuuren, Genade S, Lochner A (2010) Kinases and phosphatases in ischaemic preconditioning: a re-evaluation. Basic Res Cardiol 105:495–511PubMedCrossRefGoogle Scholar
  17. Flamigni F, Stanic’ I, Facchini A, Cetrullo S, Tantini B, Borzì RM et al (2007) Polyamine biosynthesis as a target to inhibit apoptosis of non-tumoral cells. Amino Acids 33:197–202PubMedCrossRefGoogle Scholar
  18. Flamigni F, Pignatti C, Muscari C, Giordano E, Tantini B, Stefanelli C (2009) Experimental studies on the involvement of polyamines in cardiac remodeling. In: Toninello A et al (eds) Biologically active amines and related enzymes: biochemical physiological and clinical aspects. Transworld Research Network, Kerala, pp 71–83Google Scholar
  19. Fu YC, Chi CS, Yin SC, Hwang B, Chiu YT, Hsu SL (2004a) Norepinephrine induces apoptosis in neonatal rat cardiomyocytes through a reactive oxygen species-TNF alpha-caspase signaling pathway. Cardiovasc Res 62:558–567PubMedCrossRefGoogle Scholar
  20. Fu YC, Chi CS, Yin SC, Hwang B, Chiu YT, Hsu SL (2004b) Norepinephrine induces apoptosis in neonatal rat endothelial cells via down-regulation of Bcl-2 and activation of beta-adrenergic and caspase-2 pathways. Cardiovasc Res 61:143–151PubMedCrossRefGoogle Scholar
  21. Griendling KK, Sorescu D, Lassègue B, Ushio-Fukai M (2000) Modulation of protein kinase activity and gene expression by reactive oxygen species and their role in vascular physiology and pathophysiology. Arterioscler Thromb Vasc Biol 20(10):2175–2183PubMedGoogle Scholar
  22. Gupta MK, Neelakantan TV, Sanghamitra M, Tyagi RK, Dinda A, Maulik S et al (2006) An assessment of the role of reactive oxygen species and redox signaling in norepinephrine-induced apoptosis and hypertrophy of H9c2 cardiac myoblasts. Antioxid Redox Signal 8:1081–1093PubMedCrossRefGoogle Scholar
  23. Haft JI (1974) Cardiovascular injury induced by sympathetic catecholamines. Prog Cardiovasc Dis 17:73–86PubMedCrossRefGoogle Scholar
  24. Hausenloy DJ, Tsang A, Mocanu MM, Yellon DM (2005) Ischemic preconditioning protects by activating prosurvival kinases at reperfusion. Am J Physiol Heart Circ Physiol 288:H971–H976PubMedCrossRefGoogle Scholar
  25. Igata M, Motoshima H, Tsuruzoe K, Kojima K, Matsumura T, Kondo T et al (2005) Adenosine monophosphate-activated protein kinase suppresses vascular smooth muscle cell proliferation through the inhibition of cell cycle progression. Circ Res 97:837–844PubMedCrossRefGoogle Scholar
  26. Ivaska J, Nissinen L, Immonen N, Eriksson JE, Kähäri VM, Heino J (2002) Integrin α2 β1 promotes activation of protein phosphatase 2A and dephosphorylation of Akt and glycogen synthase kinase 3 β. Mol Cell Biol 22:1352–1359PubMedCrossRefGoogle Scholar
  27. Kefas BA, Cai Y, Ling Z, Heimberg H, Hue L, Pipeleers D et al (2003) AMP-activated protein kinase can induce apoptosis of insulin-producing MIN6 cells through stimulation of c-Jun-N-terminal kinase. J Mol Endocrinol 30:151–161PubMedCrossRefGoogle Scholar
  28. Kniewald H, Malcic I, Radosevic K, Gaurina Srcek V, Slivac I, Polancec D, Matijasic M, Kniewald J, Kniewald Z (2007) Application of flow cytometry in the study of apoptosis in neonatal rat cardiomyocytes. Methods Find Exp Clin Pharmacol 29(10):681–687PubMedCrossRefGoogle Scholar
  29. Levkau B, Schäfers M, Wohlschlaeger J, von Wnuck Lipinski K, Keul P, Hermann S et al (2008) Survivin determines cardiac function by controlling total cardiomyocyte number. Circulation 117:1583–1593PubMedCrossRefGoogle Scholar
  30. Liu W, Akhand AA, Takeda K, Kawamoto Y, Itoigawa M, Kato M et al (2003) Protein phosphatase 2A-linked and -unlinked caspase-dependent pathways for downregulation of Akt kinase triggered by 4-hydroxynonenal. Cell Death Differ 10:772–781PubMedCrossRefGoogle Scholar
  31. Mao W, Iwai C, Keng PC, Vulapalli R, Liang CS (2006) Norepinephrine-induced oxidative stress causes PC-12 cell apoptosis by both endoplasmic reticulum stress and mitochondrial intrinsic pathway: inhibition of phosphatidylinositol 3-kinase survival pathway. Am J Physiol Cell Physiol 290(5):C1373–C1384PubMedCrossRefGoogle Scholar
  32. Meisse D, Van de Casteele M, Beauloye C, Hainault I, Kefas BA, Rider MH et al (2002) Sustained activation of AMP-activated protein kinase induces c-Jun N-terminal kinase activation and apoptosis in liver cells. FEBS Lett 526:38–42PubMedCrossRefGoogle Scholar
  33. Ni YG, Wang N, Cao DJ, Sachan N, Morris DJ, Gerard RD et al (2007) FoxO transcription factors activate Akt and attenuate insulin signaling in heart by inhibiting protein phosphatases. Proc Natl Acad Sci USA 104:20517–20522PubMedCrossRefGoogle Scholar
  34. Pignatti C, Tantini B, Stefanelli C, Flamigni F (2004) Signal transduction pathways linking polyamines to apoptosis. Amino Acids 27:359–365PubMedCrossRefGoogle Scholar
  35. Ray RM, Viar MJ, Yuan Q, Johnson IR (2000) Polyamine depletion delays apoptosis of rat intestinal epithelial cells. Am J Physiol Cell Physiol 278:C480–C489PubMedGoogle Scholar
  36. Ray RM, Bhattacharya S, Johnson LR (2005) Protein phosphatase 2A regulates apoptosis in intestinal epithelial cells. J Biol Chem 280:31091–31100PubMedCrossRefGoogle Scholar
  37. Song JQ, Teng X, Cai Y, Tang CS, Qi YF (2009) Activation of Akt/GSK-3b signaling pathway is involved in intermedin1-53 protection against myocardial apoptosis induced by ischemia/reperfusion. Apoptosis 14:1299–1307PubMedCrossRefGoogle Scholar
  38. Sontag E (2001) Protein phosphatase 2A: the trojan horse of cellular signaling. Cell Signal 13:7–16PubMedCrossRefGoogle Scholar
  39. Stanic’ I, Facchini A, Borzì RM, Stefanelli C, Flamigni F (2009) The polyamine analogue N1, N11-diethylnorspermine can induce chondrocyte apoptosis independently of its ability to alter metabolism and levels of natural polyamines. J Cell Physiol 219:109–116PubMedCrossRefGoogle Scholar
  40. Stanic′ I, Facchini A, Borzì RM, Vitellozzi R, Stefanelli C, Goldring MB et al (2006) Polyamine depletion inhibits apoptosis following blocking of survival pathways in human chondrocytes stimulated by tumor necrosis factor-alpha. J Cell Physiol 206:138–146CrossRefGoogle Scholar
  41. Stefanelli C, Pignatti C, Tantini B, Fattori M, Stanic I, Mackintosh CA et al (2001) Effect of polyamine depletion on caspase activation: a study with spermine synthase-deficient cells. Biochem J 355:199–206PubMedCrossRefGoogle Scholar
  42. Sugden PH, Clerk A (1998) Cellular mechanisms of cardiac hypertrophy. J Mol Med 76:725–746PubMedCrossRefGoogle Scholar
  43. Tantini B, Fiumana E, Cetrullo S, Pignatti C, Bonavita F, Shantz LM et al (2006) Involvement of polyamines in apoptosis of cardiac myoblasts in a model of simulated ischemia. J Mol Cell Cardiol 40:775–782PubMedCrossRefGoogle Scholar
  44. Tipnis UR, He GY, Li S, Campbell G, Boor PJ (2000) Attenuation of isoproterenol-mediated myocardial injury in rat by an inhibitor of polyamine synthesis. Cardiovasc Pathol 9:273–280PubMedCrossRefGoogle Scholar
  45. Tung HY, Pelech S, Fisher MJ, Pogson CI, Cohen P (1985) The protein phosphatases involved in cellular regulation. Influence of polyamines on the activities of protein phosphatase-1 and protein phosphatase-2A. Eur J Biochem 149:305–313PubMedCrossRefGoogle Scholar
  46. Vanhaesebroeck B, Alessi DR (2000) The PI3K-PDK1 connection: more than just a road to PKB. Biochem J 346:561–576PubMedCrossRefGoogle Scholar
  47. Wang Y, Huang S, Sah VP, Ross J Jr, Brown JH, Han J et al (1998a) Cardiac muscle cell hypertrophy and apoptosis induced by distinct members of the p38 mitogen-activated protein kinase family. J Biol Chem 273:2161–2168PubMedCrossRefGoogle Scholar
  48. Wang L, Ma W, Markovich R, Chen JW, Wang PH (1998b) Regulation of cardiomyocyte apoptotic signaling by insulin-like growth factor I. Circ Res 83:516–522PubMedGoogle Scholar
  49. Yasuoka C, Ihara Y, Ikeda S, Miyahara Y, Kondo T, Kohono S (2004) Antiapoptotic activity of Akt is down-regulated by Ca2+ in myocardiac H9c2 cells. J Biol Chem 279:51182–51192PubMedCrossRefGoogle Scholar
  50. Zaugg M, Jamali NZ, Lucchinetti E, Shafiq SA, Siddiqui MA (2000) Norepinephrine-induced apoptosis is inhibited in adult rat ventricular myocytes exposed to volatile anesthetics. Anesthesiology 93(1):209–218PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Silvia Cetrullo
    • 1
  • Benedetta Tantini
    • 1
    Email author
  • Annalisa Facchini
    • 1
  • Carla Pignatti
    • 1
  • Claudio Stefanelli
    • 1
  • Claudio Marcello Caldarera
    • 2
  • Flavio Flamigni
    • 1
  1. 1.Department of Biochemistry “G. Moruzzi”University of BolognaBolognaItaly
  2. 2.National Institute for Cardiovascular ResearchBolognaItaly

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