Advertisement

Cardiovascular Drugs and Therapy

, Volume 21, Issue 4, pp 227–233 | Cite as

Necrostatin: A Potentially Novel Cardioprotective Agent?

  • Christopher C. T. Smith
  • Sean M. Davidson
  • Shiang Y. Lim
  • James C. Simpkin
  • John S. Hothersall
  • Derek M. YellonEmail author
Article

Abstract

Background

Necrostatin-1 (Nec-1), a small tryptophan-based molecule, was recently reported to protect the cerebral cortex against ischemia-reperfusion (I/R) injury. We investigated the actions of Nec-1 and its so-called inactive analog, Nec-1i, in the setting of myocardial I/R injury.

Materials and methods

The actions of Nec-1 and Nec-1i were examined in cultured C2C12 and H9c2 myocytes, cardiomyocytes isolated from male Sprague–Dawley rats, Langendorff isolated perfused C57Bl/6J mouse hearts and an in vivo open-chest C57Bl/6J mouse heart model.

Results

Nec-1 at 30 μM and 100 μM (but not 100 μM Nec-1i) reduced peroxide-induced cell death in C2C12 cells from 51.2 ± 1.1% (control) to 26.3 ± 2.9% (p < 0.01 vs control) and 17.8 ± 0.9% (p < 0.001), respectively. With H9c2 cells cell death was also reduced from 73.0 ± 0.4% (control) to 56.7 ± 0% (30 μM Nec-1, p < 0.05) and 45.4 ± 3.3% (100 μM Nec-1, p < 0.01). In the isolated perfused heart Nec-1 (30 μM) reduced infarct size (calculated as a percentage of the risk area) from 48.0 ± 2.0% (control) to 32.1 ± 5.4% (p < 0.05). Nec-1i (30 μM) also reduced infarct size (32.9 ± 5.1%, p < 0.05). In anesthetized C57Bl/6J mice Nec-1 (1.65 mg/kg), given intraperitoneally to coincide with reperfusion following left anterior descending artery ligation (30 min), also reduced infarct size from 45.3 ± 5.1% (control) to 26.6 ± 4.0% (p < 0.05), whilst Nec-1i (1.74 mg/kg) was ineffective (37.8 ± 6.0%). Stimulus-induced opening of the mitochondrial permeability transition pore (MPTP) in rat cardiomyocytes, as reflected by the time until mitochondrial depolarisation, was unaffected by Nec-1 or Nec-1i at 30 μM but increased at 100 μM i.e. 91% (p < 0.05 vs control) and 152% (p < 0.001) for Nec-1 and Nec-1i, respectively.

Conclusion

This is the first study to demonstrate that necrostatins inhibit myocardial cell death and reduce infarct size, possibly via a mechanism independent of the MPTP.

Key words

necrostatin cardioprotection infarct size mitochondrial permeability transition pore 

Notes

Acknowledgements

This project was supported by a Programme Grant from the British Heart Foundation.

References

  1. 1.
    Freude B, Masters TN, Kostin S, Robicsek F, Schaper J. Cardiomyocyte apoptosis in acute and chronic conditions. Basic Res Cardiol 1998;93:85–9.PubMedCrossRefGoogle Scholar
  2. 2.
    Halestrap AP, Clarke SJ, Javadov SA. Mitochondrial permeability transition pore opening during myocardial reperfusion—a target for cardioprotection. Cardiovasc Res 2004;61:372–85.PubMedCrossRefGoogle Scholar
  3. 3.
    Hausenloy DJ, Duchen MR, Yellon DM. Inhibiting mitochondrial permeability transition pore opening at reperfusion protects against ischaemia-reperfusion injury. Cardiovasc Res 2003;63:305–12.CrossRefGoogle Scholar
  4. 4.
    Hausenloy DJ, Yellon DM. New directions for protecting the heart against ischaemia-reperfusion injury: targeting the reperfusion injury salvage kinase (RISK)-pathway. Cardiovasc Res 2004;61:448–60.PubMedCrossRefGoogle Scholar
  5. 5.
    Efthymiou CA, Mocanu MM, Yellon DM. Atorvastatin and myocardial reperfusion injury: new pleiotropic effect implicating multiple prosurvival signaling. J Cardiovasc Pharmacol 2005;45:247–52.PubMedCrossRefGoogle Scholar
  6. 6.
    Wynne AM, Mocanu MM, Yellon DM. Pioglitazone mimics preconditioning in the isolated perfused rat heart: a role for the prosurvival kinases PI3K and p42/44 MAPK. J Cardiovasc Pharmacol 2005;46:817–22.PubMedCrossRefGoogle Scholar
  7. 7.
    Baxter GF, Ebrahim Z. Role of bradykinin in preconditioning and protection of the ischaemic myocardium. Br J Pharmacol 2002;135:843–54.PubMedCrossRefGoogle Scholar
  8. 8.
    Jonassen AK, Sack MN, Mjos OD, Yellon DM. Myocardial protection by insulin at reperfusion requires early administration and is mediated via Akt and p70s6 kinase cell-survival signaling. Circ Res 2001;89:1191–8.PubMedGoogle Scholar
  9. 9.
    Bullard AJ, Govewalla P, Yellon DM. Erythropoietin protects the myocardium against reperfusion injury in vitro and in vivo. Basic Res Cardiol 2005;100:397–403.PubMedCrossRefGoogle Scholar
  10. 10.
    Bose AK, Mocanu MM, Carr RD, Brand CL, Yellon DM. Glucagon-like peptide 1 can directly protect the heart against ischemia/reperfusion injury. Diabetes 2005;54:146–51.PubMedCrossRefGoogle Scholar
  11. 11.
    Smith CCT, Mocanu MM, Davidson SM, Wynne AM, Simpkin JC, Yellon DM. Leptin, the obesity-associated hormone, exhibits direct cardioprotective activity. Br J Pharmacol 2006;149:5–13.PubMedCrossRefGoogle Scholar
  12. 12.
    Degterev A, Huang Z, Boyce M, Li Y, Jagtap P, Mizushima N, et al. Chemical inhibitor of non apoptotic cell death with therapeutic potential for ischemic brain injury. Nature Chem Biol 2005;1:112–9.CrossRefGoogle Scholar
  13. 13.
    Bao L, Li Y, Deng S-X, Landry D, Tabas I. Sitosterol-containing lipoproteins trigger free sterol-induced caspase-independent death in ACAT-competent macrophages. J Biol Chem 2006;281:33635–49.PubMedCrossRefGoogle Scholar
  14. 14.
    Kukan M. Emerging roles of proteasomes in ischemia-reperfusion injury of organs. J Physiol Pharmacol 2004;55:3–15.PubMedGoogle Scholar
  15. 15.
    Hausenloy DJ, Yellon DM, Mani-Babu S, Duchen MR. Preconditioning protects by inhibiting the mitochondrial permeability transition. Am J Physiol Heart Circ Physiol 2004;287:H841–9.PubMedCrossRefGoogle Scholar
  16. 16.
    Jacobson J, Duchen MR. Mitochondrial oxidative stress and cell death in astrocytes—requirement for stored Ca2+ and sustained opening of the permeability transition pore. J Cell Sci 2002;115:1175–88.PubMedGoogle Scholar
  17. 17.
    Sumeray MS, Yellon DM. Characterisation and validation of a murine model of global ischaemia-reperfusion injury. Mol Cell Biochem 1998;186:61–8.PubMedCrossRefGoogle Scholar
  18. 18.
    Marber MS, Mestril R, Chi SH, Sayen MR, Yellon DM, Dillman WH. Overexpression of the rat inducible 70-kD heat stress protein in a transgenic mouse increases the resistance of the heart to ischaemic injury. J Clin Invest 1995;95:1446–56.PubMedCrossRefGoogle Scholar
  19. 19.
    Lim SY, Wainwright CL, Kennedy S, Kane KA. Activation of protease-activated receptor-2 (PAR-2) induces cardioprotection in anaesthetised mice. J Mol Cell Cardiol 2005;38:858.CrossRefGoogle Scholar
  20. 20.
    Formigli L, Papucci L, Tani A, Schiavone N, Tempestini A, Orlandini GE, et al. Aponecrosis: morphological and biochemical exploration of a syncretic process of cell death sharing apoptosis and necrosis. J Cell Physiol 2000;182:41–9.PubMedCrossRefGoogle Scholar
  21. 21.
    Mocanu MM, Baxter GF, Yellon DM. Caspase inhibition and limitation of myocardial infarct size: protection against lethal reperfusion injury. Br J Pharmacol 2000;130:197–200.PubMedCrossRefGoogle Scholar
  22. 22.
    Teng X, Degterev A, Jagatap P, Xing X, Choi S, Denu R, et al. Structure-activity relationship study of novel necroptosis inhibitors. Bioorg Med Chem Lett 2005;15:5039–44.PubMedCrossRefGoogle Scholar
  23. 23.
    Zhao ZQ, Vinten-Johansen J. Myocardial apoptosis and ischemic preconditioning. Cardiovasc Res 2002;55:438–55.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2007

Authors and Affiliations

  • Christopher C. T. Smith
    • 1
  • Sean M. Davidson
    • 1
  • Shiang Y. Lim
    • 1
  • James C. Simpkin
    • 1
  • John S. Hothersall
    • 1
  • Derek M. Yellon
    • 1
    Email author
  1. 1.The Hatter Cardiovascular InstituteUniversity College London Hospital and Medical SchoolLondonUK

Personalised recommendations