Abstract
Hypothermic perfusion of rat livers was investigated by 31phosphorus nuclear magnetic resonance (31P NMR) spectroscopy using a temperature-controlled module that allowed data acquisition at various time points during a 48-h period. The livers were perfused with an oxygenated lactobionate/raffinose-based solution containing adenosine and inorganic phosphate, and changes in tissue oedema were monitored by direct on-line measurements of liver weight changes. Liver tissue ATP concentrations, determined by fluorimetric assay, were low immediately after organ removal, probably reflecting metabolic stress during the removal period, and these increased slightly during the next 3 h. This was reflected by changes in the 31P NMR spectra. However, by 24 h ATP levels had increased significantly, and these were maintained for up to 48 h, suggesting a shift in the balance between energy production and consumption. When inorganic phosphate was replaced by another anion (citrate), ATP was maintained at a constant lower level during perfusion for 48 h. Tissue weight changes were similar in both groups, suggesting that volume control was not affected by the different ATP contents of the livers. By combining the temperature-controlled module with a separate perfusion circuit, NMR spectroscopy can provide a sensitive method for following energy metabolism in the same organ over long periods during hypothermic perfusion.
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References
Attenburrow V, Fuller BJ, Hobbs KEF (1981) Effects of temperature and methods of hypothermic preservation on hepatic energy metabolism. Cryo-Lett 2:15–20
Calne RY, Dunn DC, Herbertson BM, Gordon EM, Bitter-Suermann H, Robson AJ, McDonald A, Davis D, Smith D, Reitter F, Webster L (1972) Liver preservation by single passage hypothermic ‘squirt perfusion’. BMJ 4:142–148
Colman JR, Britton R, Orrego H, Saldiva V, Medline A, Israel Y (1983) Relation between osmotically induced hepatocyte enlargement and portal hypertension. Am J Physiol 245:G382-G386
D'Alessandro A, Southard JH, Kalayoglu M, Belzer FO (1986) Comparison of cold storage and perfusion of dog livers on function of tissue slices. Cryobiology 23:161–167
Delmas-Beauvieux M-C, Gallis J-L, Clerc M, Cannioni P (1992) A nuclear magnetic resonance study of thermal transition during reperfusion of rat liver preserved in Bretschneider's Solution. Transplantation 53:709–711
Delmas-Beauvieux M-C, Gallis J-L, Rousse N, Clerc M, Canioni P (1992) Phosphorus-31 nuclear magnetic resonance of isolated rat liver during hypothermic ischaemia and subsequent normothermic perfusion. J Hepatol 15:192–201
Fuller BJ, Attenburrow V, Newsholme C (1978) Experimental studies on continuous hypothermic liver perfusion with a synthetic solution containing gelatin polypeptides (Haemaccel). Cryobiology 15:279–289
Fuller BJ, Busza AL, Proctor E, Myles M, Gadian DG, Hobbs KEF (1988) Control of pH during hypothermic liver storage: role of the storage solution. Transplantation 45:239–241
Fuller BJ, Busza AL, Proctor E (1990) Possible resuscitation of liver function by hypothermic reperfusion in vitro after prolonged (24-hour) preservation —a 31P NMR study. Transplantation 50:511–513
Gulik TM van, Lindell SL, Boudjema K, Pienaar BH, Vreugdenhil PK, Southard JH, Belzer FO (1990) Combined cold storage and perfusion preservation of the canine liver. Transplant Proc 22:520–522
Jeremy JY, Stansby G, Fuller B, Rolles K, Hamilton G (1992) The effect of cold-storage of rat thoracic aortic rings in organ preservation solutions — a study of receptor-linked vascular prostacyclin synthesis. Transplantation 53:999–1002
Lockett CJ, Proctor E, Fuller BJ, Busza AL (1991) A perfusion system for studying organ preservation by nuclear magnetic resonance spectroscopy. Cryo-Lett 12:207–214
Lowry O, Passonneau H (1972) A flexible system of enzymatic analysis. Academic Press, New York
Lund P, Cornell N, Krebs H (1975) Effect of adenosine on the adenine nucleotide content and metabolism of hepatocytes. Biochem J 50:511–514
Matsuno N, Kozaki M, Sakurai E, Uchiyama M, Iwahori T, Kozaki K, Kono K, Tanaka M, Tamaki T, Tamaki I (1993) Effect of combination in situ cooling and machine perfusion preservation on non-heart-beating donor kidney procurement. Transplant Proc 25:1516–1517
McAnulty JF, Southard JH, Belzer FO (1987) Improved maintenance of adenosine-triphosphate in 5-day perfused kidneys with adenine and ribose. Transplant Proc 19:1376–1379
McAnulty JF, Vreugdenhil P, Southard J, Belzer FO (1991) Effect of a protease inhibitor on dog kidneys preserved for 5 days (5°C). Cryobiology 28:517
Pienaar BH, Lindell SL, Gulik T van, Southard JH, Belzer FO (1990) Seventy-two-hour preservation of the canine liver by machine perfusion. Transplantation 49:258–260
Yamamoto N, Konishi Y, Wakashiro S, Takayasu T, Tatsumi Y, Shimahara Y, Tanaka K, Yamaoka Y, Ozawa K (1991) 72-hour preservation of porcine liver by continuous hypothermic perfusion with UW solution in comparison with simple cold-storage. J Surg Res 51:288–292
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Lockett, C.J., Fuller, B.J., Busza, A.L. et al. Hypothermic perfusion preservation of liver: the role of phosphate in stimulating ATP synthesis studied by 31P NMR. Transpl Int 8, 440–445 (1995). https://doi.org/10.1007/BF00335595
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DOI: https://doi.org/10.1007/BF00335595