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31p-NMR spectroscopy of rat liver during simple storage or continuous hypothermic perfusion

Abstract

The ATP content and intracellular pH (pH(i)) of isolated rat liver before, during, and after cold preservation in either UW-lactobionate (UW, n=10) or Euro-Collins (EC, n=8) solutions were monitored using phosphorus-31 nuclear magnetic resonance ((31)P-NMR) spectroscopy. The (31)P-NMR spectra were obtained on a 4.7-Tesla system operating at 81 MHz. Fructose metabolism, liver enzyme release, O(2) consumption, and rat survival after liver transplantation were also evaluated. During simple cold storage (SCS), the ATP level declined to undetectable levels with both preservation solutions while the pH(i) declined to approximately 7.0. In contrast, during continuous hypothermic perfusion (CHP), hepatic ATP levels remained measurable during the 24-hour EC preservation and actually increased significantly (p>0.01) during UW preservation. After reperfusion at 37°C with Krebs-lactate, the SCS livers treated with EC differed significantly from the UW livers in terms of their ATP and pH(i) as well as their response to a fructose challenge. In contrast, livers undergoing CHP demonstrated similar behaviors with both solutions. These results demonstrate an increase in the hepatic ATP content during CHP which occurs with UW but is not seen with EC. On the other hand, only livers that were simply stored with UW achieved significant survival after transplant, while CHP livers were affected by vascular damage as demonstrated by fatal thrombosis after transplant. These data suggest that ATP content is not the only determinant of good liver function although a system of hypothermic perfusion might further improve liver preservation efficacy should injury to vascular endothelium be avoided

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