Basic Research in Cardiology

, Volume 90, Issue 5, pp 388–396 | Cite as

Substrate selection in the isolated working rat heart: effects of reperfusion, afterload, and concentration

  • F. M. H. Jeffrey
  • V. Diczku
  • A. D. Sherry
  • C. R. Malloy
Original Contribution


A study of substrate selection in the isolated heart was made using13C NMR isotopomer analysis, a method that unequivocally identifies relative substrate utilization. This technique has several advantages over conventional approaches used to study this problem. It detects the labeling of metabolic end-products present in tissue, as opposed to more indirect methods such as measurement of respiratory quotient, arteriovenous differences, or specific activity changes in the added substrate. It also has advantages over methods such as14CO2 release, which may involve dilution of label with unlabeled pools before CO2 release. Furthermore, it can measure the relative oxidation of up to four substrates in a single experiment, which other labeling techniques cannot conveniently achieve. Substrates selection was considered in light of its effects on myocardial efficiency and recovery from ischemia. A mixture of four substrates (acetoacetate, glucose, lactate, and a mixture of long chain fatty acids), present at physiological concentration (0.17, 5.5, 1.2, and 0.35 mM, respectively), was examined. This is the first use of such a mixture in the study of substrate selection in an isolated organ preparation. At these concentrations, it was found that fatty acids supplied the majority of the acetyl-CoA (49%), and a substantial contribution was also provided by acetoacetate (23%). This suggests that the ketone bodies are a more important substrate than generally considered. Indeed, normalizing the relative utilizations on the basis of acetyl-CoA equivalents, ketone bodies were by far the preferred substrate. The relative lactate oxidation was only 15%, and glucose oxidation could not be detected. No change in utilization was detected after 15 min of ischemia followed by 40 min of reperfusion. The change in substrate selection with afterload was examined, to mimic the stress-related changes in workload found with ischemia. Only minor changes were found. Substrate selection from the same group of substrates, but employing concentrations observed during starvation, was also assessed. This represents the state during which most clinical treatments and evaluations are performed. In this case, acetoacetate was the most used substrate (78%), with small and equal contributions from fatty acids and endogenous substrates; the oxidation of lactate was suppressed.

Key words

Substrate selection isolated rat heart 13C NMR 


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  1. 1.
    Bing RJ (1965) Cardiac Metabolism. Physiol Rev 45: 171–213Google Scholar
  2. 2.
    Bing RJ, Siegel A, Ungar I, Gilbert M (1954) Metabolism of the human heart. II. Studies on fat, ketone and amino acid metabolism. Am J Med 16: 504–515Google Scholar
  3. 3.
    Bing RJ, Siegel A, Vitale A, Balboni F, Sparks E, Taeschler M, Klapper M, Edwards S (1953) Metabolic studies on the human heart in vivo. I. Studies on carbohydrate metabolism of the human heart. Am J Med 15: 284–296Google Scholar
  4. 4.
    Bowman RH (1962) The effect of longchain fatty acids on glucose utilization in the isolated perfused rat heart. Biochem J 84: 14pGoogle Scholar
  5. 5.
    Challoner DR, Steinberg D (1966) Effect of free fatty acid on the oxygen consumption of perfused rat heart. Am J Physiol 210: 280–6Google Scholar
  6. 6.
    Crass MF 3d, McCaskill ES, Shipp JC (1969) Effect of pressure development on glucose and palmitate metabolism in persused heart. Am J Physiol 216: 1569–76Google Scholar
  7. 7.
    Drake AJ (1982) Substrate utilization in the myocardium. Basic Res Cardiol 77: 76Google Scholar
  8. 8.
    Drake AJ, Haines JR, Noble MI (1980) Preferential uptake of lactate by the normal myocardium in dogs. Cardiovasc Res 14: 65–72Google Scholar
  9. 9.
    Drake-Holland AJ, Elzinga G, Noble MI, ter Keurs HE, Wempe FN (1983) The effect of palmitate and lactate on mechanical performance and metabolism of cat and rat myocardium. J Physiol (Lond) 339: 1–15Google Scholar
  10. 10.
    Garland PB, Newsholme EA, Randle PJ (1962) Effects of fatty acids, ketone bodies, diabetes and starvation on pyruvate metabolism in rat heart and diaphragm muscle. Nature 195: 381Google Scholar
  11. 11.
    Gmeiner R, Apstein CS, Brachfeld N (1975) Effect of palmitate on hypoxic cardiac performance. J Mol Cell Cardiol 7: 227–35Google Scholar
  12. 12.
    Gorge G, Chatelain P, Schaper J, Lerch R (1991) Effect of increasing degrees of ischemic injury on myocardial oxidative metabolism early after reperfusion in isolated rat hearts. Circ Res 68: 1681–92Google Scholar
  13. 13.
    Hall LM (1961) Preferential oxidation of acetoacetate by the perfused heart. Biochem Biophys Res Commun 6: 177–179Google Scholar
  14. 14.
    Issekutz B Jr, Miller HI, Paul P, Podahl K (1965) Effect of lactic acid on free fatty acids and glucose oxidation in dogs. Am J Physiol 209: 1137–44Google Scholar
  15. 15.
    Jeffrey FM, Rajagopal A, Malloy CR, Sherry AD (1991) 13C-NMR: a simple yet comprehensive method for analysis of intermediary metabolism. Trends Biochem Sci 16: 5–10Google Scholar
  16. 16.
    Kahles H, Hellige G, Hunneman DH, Mezger VA, Bretschneider HJ (1982) Influence of myocardial substrate utilization on the oxygen consumption of the heart. Clin Cardiol 5: 286–93Google Scholar
  17. 17.
    Kahles H, Schafer W, Lick T, Junggeburth J, Kochsiek K (1986) Changes in myocardial substrate and energy metabolism by S-(4)-hydroxyphenyl-glycine and an N-(6)-derivative of adenosine. Basic Res Cardiol 81: 258–66Google Scholar
  18. 18.
    Kiviluoma KT, Karhunen M, Lapinlampi T, Peuhkurinen KJ, Hassinen IE (1988) Acetate-induced changes in cardiac energy metabolism and hemodynamics in the rat. Basic Res Cardiol 83: 431–44Google Scholar
  19. 19.
    Liedtke AJ, DeMaison L, Eggleston AM, Cohen LM, Nellis SH (1988) Changes in substrate metabolism and effects of excess fatty acids in reperfused myocardium. Circ Res 62: 535–42Google Scholar
  20. 20.
    Little JR, Goto M, Spitzer JJ (1970) Effect of ketones on metabolism of FFA by dog myocardium and skeletal muscle in vivo. Am J Physiol 219: 1458–63Google Scholar
  21. 21.
    Lopaschuk GD, Spafford MA, Davies NJ, Wall SR (1990) Glucose and palmitate oxidation in isolated working rat hearts reperfused after a period of transient global ischemia. Circ Res 66: 546–53Google Scholar
  22. 22.
    Malloy CR, Sherry AD, Jeffrey FM (1988) Evaluation of carbon flux and substrate selection through alternate pathways involving the citric acid cycle of the heart by 13C NMR spectroscopy. J Biol Chem 263: 6964–71Google Scholar
  23. 23.
    Malloy CR, Sherry AD, Jeffrey FM (1990) Analysis of tricarboxylic acid cycle of the heart using 13C isotope isomers. Am J Physiol 259: H987–95Google Scholar
  24. 24.
    Malloy CR, Thompson JR, Jeffrey FM, Sherry AD (1990) Contribution of exogenous substrates to acetyl coenzyme A: measurement by 13C NMR under non-steady-state conditions. Biochemistry 29: 6756–61Google Scholar
  25. 25.
    Mjos OD (1971) Effect of free fatty acids on myocardial function and oxygen consumption in intact dogs. J Clin Invest 50: 1386–9Google Scholar
  26. 26.
    Most AS, Lipsky MH, Szydlik PA, Bruno C (1973) Failure of free fatty acids to influence myocardial oxygen consumption in the intact, anesthetized dog. Cardiology 58: 220–8Google Scholar
  27. 27.
    Myears DW, Sobel BE, Bergmann SR (1987) Substrate use in ischemic and reperfused canine myocardium: quantitative considerations. Am J Physiol 253: H107–14Google Scholar
  28. 28.
    Neely JR, Morgan HE (1974) Relationship between carbohydrate and lipid metabolism and the energy balance of heart muscle. Annu Rev Physiol 36: 413–59Google Scholar
  29. 29.
    Neely JR, Rovetto MJ, Oram JF (1972) Myocardial utilization of carbohydrate and lipids. Prog Cardiovasc Dis 15: 280–329Google Scholar
  30. 30.
    Olson RE (1962) Effect of pyruvate and acetoacetate on the metabolism of fatty acids by the perfused rat heart. Nature 195: 597–99Google Scholar
  31. 31.
    Randle PJ, England PJ, Denton RM (1970) control of the tricarboxytate cycle and its interactions with glycolysis during acetate utilization in rat heart. Biochem J 117: 677–95Google Scholar
  32. 32.
    Regan TJ, Binak K, Gordon S, Defazio V, Hellems HK (1961) Myocardial oxygen consumption during postprandial lipema and heparin induced lipolysis. Circulation 23: 55–63Google Scholar
  33. 33.
    Remesy C, Demigne C (1983) Changes in availability of glucogenic and ketogenic substrates and liver metabolism in fed or starved rats. Ann Nutr Metab 27: 57–70Google Scholar
  34. 34.
    Rogers WJ, McDaniel HG, Moraski RE, Rackley CE, Russell RO Jr (1977) Effect of heparin-induced free fatty acid elevation on myocardial oxygen consumption in man. Am J Cardiol 40: 365–72Google Scholar
  35. 35.
    Saddik M, Lopaschuk GD (1991) Myocardial triglyceride turnover and contribution to energy substrate utilization in isolated working rat hearts. J Biol Chem 266: 8162–70Google Scholar
  36. 36.
    Sherry AD, Sumegi B, Miller B, Cottam GL, Gavva S, Jones JG, Malloy CR (1994) Orientation-conserved transfer of symmetric Krebs cycle intermediates in mammalian tissue. Biochemistry 33: 6268–75Google Scholar
  37. 37.
    Solomon MA, Jeffrey FMH, Storey CJ, Sherry AD, Malloy CR (1993) 13C Isotopomer Analysis of Myocardial Substrate Selection in Early Reperfusion After 15 Minutes of Regional Ischemia in the Working Rabbit Heart. Soc Mag Reson Med 2: 1052Google Scholar
  38. 38.
    Spitzer JJ (1974) Effect of lactate infusion on canine myocardial free fatty acid metabolism in vivo. Am J Physiol 226: 213–7Google Scholar
  39. 39.
    Spitzer JJ, Spitzer JA (1972) Myocardial metabolism in dogs during hemorrhagic shock. Am J Physiol 222: 101–5Google Scholar
  40. 40.
    Taegtmeyer H, Hems R, Krebs HA (1980) Utilization of energy-providing substrates in the isolated working rat heart. Biochem J 186: 701–11Google Scholar
  41. 41.
    van der Vusse GJ, Glatz JF, Stam HC, Reneman RS (1992) Fatty acid homeostasis in the normoxic and ischemic heart. Physiol Rev 72: 881–940Google Scholar
  42. 42.
    Vik-Mo H, Mjos OD (1981) Influence of free fatty acids on myocardial oxygen consumption and ischemic injury. Am J Cardiol 48: 361–5Google Scholar
  43. 43.
    Waters ET, Fletcher JP, Mirsky IA (1938) The relation between carbohydrate and betahydroxybutyric acid utilization by the heart-lung preparation. Am J Physiol 122: 542Google Scholar
  44. 44.
    Willebrands AF, van der Veen KJ (1967) Influence of substrate on oxygen consumption of isolated perfused rat heart. Am J Physiol 212: 1529–35Google Scholar
  45. 45.
    Williamson JR, Krebs HA (1961) Acetoacetate as fuel of respiration in the perfused rat heart. Biochem J 80: 540–547Google Scholar
  46. 46.
    Zierler KL (1976) Fatty acids as substrates for heart and skeletal muscle. Circ Res 38: 459–63Google Scholar

Copyright information

© Springer-Verlag 1995

Authors and Affiliations

  • F. M. H. Jeffrey
    • 1
  • V. Diczku
    • 2
  • A. D. Sherry
    • 1
    • 2
  • C. R. Malloy
    • 1
    • 3
  1. 1.Department of Radiology UT Southwestern Medical CenterThe Mary Nell and Ralph B. Rogers Magnetic Resonance CenterDallasUSA
  2. 2.Department of ChemistryUniversity of Texas at DallasRichardsonUSA
  3. 3.Department of Internal MedicineUT Southwestern Medical CenterDallasUSA

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