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Cardiovascular Research 1997 34(3):547-556; doi:10.1016/S0008-6363(97)00058-8
© 1997 by European Society of Cardiology
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Copyright © 1997, European Society of Cardiology

Alteration in the control of mitochondrial respiration by outer mitochondrial membrane and creatine during heart preservation

Laurence Kay, Zoya Daneshrad, Valdur A. Saks1,1 and André Rossi*

Laboratoire de Bioénergétique, Université Joseph Fourier, BP 53-38041, Grenoble Cedex 9, France

* Corresponding author. Tel.: +33 04 76 51 46 70; fax: +33 04 76 51 42 18.

Objective: This study aimed to evaluate the nature and extent of mitochondrial alterations during heart preservation. Methods: Rat hearts, isolated after cardioplegia in situ, were preserved for 6 or 15 h at 4°C either by immersion in cardioplegic solution or by low-flow perfusion (0.3 ml/min) with cardioplegic solution. The energy state of hearts at the end of preservation was determined by 31P-NMR spectroscopy and functional recovery was evaluated on reperfusion. Variables of mitochondrial respiration (maximal rate of respiration in the presence of ADP={Delta}Vmax, apparent Km for ADP, effect of creatine) were evaluated on skinned fibers and compared with those determined in controls and in hearts subjected to 1 hour of ischemia at 37°C. Results: Serious mitochondrial alterations were detected in fibers from 15 h immersed hearts: decrease of {Delta}Vmax and of apparent Km for ADP, loss of the stimulatory effect of creatine, and disruption of the outer mitochondrial membrane. The extent of alterations was more accentuated in fibers from normothermic ischemic hearts, in which some damage of the inner mitochondrial membrane also occured. In fibers from hearts preserved for 6 h, no significant changes in mitochondrial variables could be detected. When the hearts were preserved under low-flow perfusion for 15 h, only the stimulatory effect of creatine on respiration was significantly decreased. The extent of the loss of the stimulatory effect of creatine paralleled the accumulation of inorganic phosphate (Pi) during preservation and the decrease in left ventricular function on reperfusion. Conclusions: Alterations related to the outer membrane and the intermembrane space are among the earliest signs of damage to mitochondrial function during heart preservation. These alterations could be attributed to the swelling of mitochondria under the effect of Pi. The determination of mitochondrial parameters in biopsy samples could allow a simple and rapid evaluation of energy-producing and transfer capacities of the myocardium.

KEYWORDS Heart preservation; Myocardial ischemia; Skinned fibers; Mitochondria, outer membrane; Creatine kinase; Rat, heart

1 On leave from Laboratory of Bioenergetics, Institute of Chemical and Biological Physics, Tallinn, Estonia.


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