Basic Research in Cardiology

, Volume 91, Issue 1, pp 79–85 | Cite as

The ability of heat stress and metabolic preconditioning to protect primary rat cardiac myocytes

  • D. V. E. Cumming
  • R. J. Heads
  • N. J. Brand
  • D. M. Yellon
  • D. S. Latchman
Original Contributions


Primary rat cardiocytes were subjected to either thermal “preconditioning” for 30 min at 43°C or 20 min metabolic “preconditioning” (10 mM deoxyglucose, 20 mM lactate, pH 6.5). Eighteen hours later cells were analysed either for hsp 70i expression or subjected to a subsequent lethal heat stress or simulated ischaemia (10 mM deoxyglucose, 20 mM lactate, 0.75 mM sodium dithionite, 12 mM potassium chloride, pH 6.5) for 2 hours and assessed for survival by trypan blue exclusion.

Hsp 70i was induced over 100 fold by thermal “preconditioning” and 30 fold by metabolic “preconditioning” (p<0.001, p<0.05), hsp 90 was induced 2.71 fold and 2.24 fold (p<0.001, p<0.001) by thermal and metabolic “preconditioning” respectively, while hsp 60 was not induced by either treatment. Preconditioned cultures had improved survival against subsequent lethal heat stress or simulated ischaemia: Thermal “preconditioning” reduced death from 69.22% to 52.46% upon subsequent “lethal” heat stress and from 49.13% to 36.66% upon subsequent “lethal” simulated ischaemia. Metabolic “preconditioning” reduced cell death from 51.29% to 33.8% against subsequent “lethal” heat stress, and from 69.09% to 55.61% upon subsequent “lethal” simulated ischaemia. A second marker of cell death, the release of lactate dehydrogenase activity into the culture media, was reduced to 65% and 60% of control values for thermally preconditioned cells subjected to “lethal” heat or “lethal” simulated ischaemia respectively. Metabolically “preconditioned” cells demonstrated lactate dehydrogenase activity of 59% and 51% that of control values, when subjected to “lethal” heat or “lethal” simulated “ischaemia” respectively.

Key words

Heat stress ischaemia hsp70 cardiac myocytes 



heat stress protein

hsp 70i

inducible 70 kDa heat stress protein


lactate dehydrogenase


phosphate buffered saline


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Amrani M, Allen NJ, O'Shea J, Corbett J, Dunn MJ, Tadjkarimi S Theodropoulos, Pepper J, Yacoub MH (1993) Role of catalase and heat shock protein on recovery of cardiac endothelial and mechanical function after ischaemia. Cardioscience 4: 193–198Google Scholar
  2. 2.
    Buja LM, Hagler HK, Parsons D, Chien K, Reynolds RC, Willerson JT (1988) Alterations of ultra structure and elemental composition in cultured neonatal rat cardiac myocytes after metabolic inhibition with iodoacetic acid. Laboratory Investigation 53: 397–412Google Scholar
  3. 3.
    Chien KR, Sen A, Reynolds R, Chang A, Kim Y, Gunn MD, Buja M, Willerson JT (1985) Release of arachidonate from membrane phospholipids in cultured neonatal rat myocardial cells during adenosine triphosphate depletion. J Clin Invest 75: 1770–1780Google Scholar
  4. 4.
    Currie RW, Karmazyn M, Kloc M, Mailer K (1988) Heat shock is associated with enhanced post ischaemic ventricular recovery. Circ Res 63: 543–549Google Scholar
  5. 5.
    Currie RW, Tanguay RM, Kingma JG (1993) Heat shock response and limitation of necrosis during occlusion/reperfusion in rabbit hearts. Circulation 87: 963–971Google Scholar
  6. 6.
    Donnely TJ, Sievers RE, Vissern FLJ, Welch WJ, Wolfe CL (1972) Heat shock protein induction in rat hearts. A role for improved myocardial salvage after ischaemia and reperfusion. Circulation 85: 769–778Google Scholar
  7. 7.
    Esumi K, Nishida M, Shaw D, Smith TW, Marsh JD (1991) NADH measurements in adult rat myocytes during simulated ischaemia. Am J Physiol 29: H1743-H1752Google Scholar
  8. 8.
    Heads RJ, Latchman DS, Yellon DM (1994) Stable high level expression of a transfected human HSP70 gene protects a heart-derived muscle cell line against thermal stress. J Molec Cell Cardiol 26: 695–699Google Scholar
  9. 9.
    Hescheler J, Meyer R, Plant S, Krautwurst D, Rosenthal W, Schultz (1991) Morphological, biochemical and electrophysiological characterisation of a clonal cell (H9c2) line from rat heart. Circ Res 69: 1476–1486Google Scholar
  10. 10.
    Hutter MM, Sievers RE, Barbosa V, Wolfe CL (1994) Heat shock protein induction in rat heart. A direct correlation between the amount of heat shock protein induced and the degree of myocardial protection. Circulation 89: 355–360Google Scholar
  11. 11.
    Iwaki K, Chi SH, Dillman WH, Mestril R (1993) Induction of HSP70 in cultured rat neonatal cardiomyocytes by hypoxia and metabolic stress. Circulation 87: 2023–2032Google Scholar
  12. 12.
    Karmazyn M, Mailer K, Currie RW (1990) Acquisition and decay of heat shock enhanced post ischaemic ventricular recovery. Am J Physiol 259: H424–431Google Scholar
  13. 13.
    Kimes BW, Brandt BL (1976) Properties of a clonal muscle cell line from rat heart. Cell Res 98: 367–381Google Scholar
  14. 14.
    Knowlton KU, Baracchini E, Ross RS, Harris AN, Henderson SA, Evans SM, Glembotski CC, Chien KR (1991) Coregulation of the atrial natriuretic factor and cardiac myosin light chain-2 genes during a adrenergic stimulation of neonatal rat ventricular cells. J Biol Chem 266: 7759–5568Google Scholar
  15. 15.
    Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of the bacteriophage T4. Nature 227: 680–685Google Scholar
  16. 16.
    Marber MS, Latchman DS, Walker JM, Yellon DM (1993) Cardiac stress protein elevation 24 hours after brief ischaemia or heat stress is associated with resistance to myocardial infarction. Circulation 88: 1264–1272Google Scholar
  17. 17.
    Marber MS, Walker JM, Latchman DS, Yellon DM (1994) Myocardial protection after whole body heat stress in the rabbit is dependent on metabolic substrate and is related to the amount of the inducible 70kD heat stress protein. J Clin Invest 93: 1087–1094Google Scholar
  18. 18.
    Mestril R, Chi SH, Sayen R, O'Reilly K, Dillmann WH (1994) Expression of inducible stress protein 80 in heart myogenic cells confers protection against simulated ischaemia-induced injury. J Clin Invest 93: 759–767Google Scholar
  19. 19.
    Murry CE, Jennings RB, Reimer KA (1986) Preconditioning with ischaemia: a delay of lethal cell injury in ischaemic myocardium. Circulation 74: 1124–1136Google Scholar
  20. 20.
    Walker DM, Pasini E, Kucukoglu S et al. (1993) Heat stress limits infarct size in the isolated perfused rabbit heart. Cardiovasc Res 27: 962–967Google Scholar
  21. 21.
    Yellon DM, Latchman DS (1992) Stress proteins and myocardiol protection. J Mol Cell Cardiol 24: 113–124Google Scholar
  22. 22.
    Yellon DM, Alkhulafi AM, Pugsley WB (1993) Preconditioning the human myocardium. The Lancet 342: 276–277Google Scholar
  23. 23.
    Yellon DM, Latchman DS, Marber MS (1993) Stress proteins: an endogenous route to cardiac protection: fact or fiction? Cardiovasc Res 27: 158–161Google Scholar
  24. 24.
    Yellon DM, Pasini E, Cargnoni A, Marber MS, Latchman DS, Ferrari R (1992) The protective role of heat stress in the ischaemic and reperfused rabbit myocardium. J Mol Cell Cardiol 24: 895–907Google Scholar

Copyright information

© Steinkopff Verlag 1996

Authors and Affiliations

  • D. V. E. Cumming
    • 1
    • 2
  • R. J. Heads
    • 1
    • 2
  • N. J. Brand
    • 3
  • D. M. Yellon
    • 1
  • D. S. Latchman
    • 2
  1. 1.The Hatter Institute for Cardiovascular Studies Department of Academic CardiologyUniversity College HospitalLondonUK
  2. 2.Medical Molecular Biology Unit Department of Molecular PathologyUniversity College London School of MedicineLondonUK
  3. 3.Department of Cardiothoracic SurgeryThe National Heart and Lung InstituteLondonUK

Personalised recommendations