The effect of dichloroacetate and alanine on the metabolic recovery of perfused mouse liver after cold ischemia.
Pyruvate dehydrogenase has been thought to be involved in the improved recovery of livers, from fasted donors, reperfused with alanine after cold preservation. The aim of this work was to investigate the effect on perfused mouse liver of dichloroacetate, an activator of this enzyme. Livers from fed and fasted animals were perfused with oxygenated Krebs-Henseleit buffer for 30 min, then stored at 4 degrees C in University of Wisconsin solution for 48 h. Then reperfusion at 37 degrees C was performed with Krebs-Henseleit buffer containing 2 mM dichloroacetate for 1 h. (3-(13)C)Alanine (8 mM) was then added and perfusion was continued for a second hour. (31)P-NMR was used to measure nucleoside triphosphate recovery of the livers. At the end of reperfusion, (13)C-NMR spectra of perfusates were recorded. Dichloroacetate (DCA) was found to activate pyruvate dehydrogenase in all cases. However, it decreased the functional recovery of livers from both fed and fasted mice. In order to study the effect of alanine on this DCA deleterious effect, we reperfused the livers according to a modified protocol. The first hour of perfusion without alanine was omitted and the organs were reperfused directly for 1 h in the presence of 2 mM dichloroacetate and 8 mM (3-(13)C)alanine. In this protocol, the deleterious effect of DCA was completely suppressed for livers from fasted mice. These results led to the conclusion that the specific beneficial effect of alanine on livers from fasted livers persists in the presence of DCA and thus cannot be explained solely by the induction of a greater pyruvate dehydrogenase reaction rate.
Patry
J
J
U350 INSERM, Institut Curie, Bâtiment 112, Centre Universitaire, F91405 Orsay Cédex, France.
Adam
R
R
Dimicoli
J L
JL
D000241:Adenosine; D000409:Alanine; D000493:Allopurinol; D000818:Animals; D002247:Carbon Isotopes; D003080:Cold Temperature; D015925:Cryopreservation / Q000379:methods*; D003999:Dichloroacetic Acid; D005215:Fasting; D005973:Glutamine / Q000378:metabolism; D005978:Glutathione; D007328:Insulin; D007511:Ischemia / Q000378:metabolism*; D008099:Liver / Q000098:blood supply* / Q000378:metabolism*; D008297:Male; D051379:Mice; D019906:Nuclear Magnetic Resonance, Biomolecular; D009926:Organ Preservation / Q000379:methods*; D019188:Organ Preservation Solutions; D010758:Phosphorus; D011887:Raffinose; D015427:Reperfusion Injury / Q000517:prevention & control; D014019:Tissue Donors
D002247:Carbon Isotopes; D007328:Insulin; D019188:Organ Preservation Solutions; C055206:University of Wisconsin-lactobionate solution; D005973:Glutamine; D010758:Phosphorus; D000493:Allopurinol; D003999:Dichloroacetic Acid; D005978:Glutathione; D000241:Adenosine; D011887:Raffinose; D000409:Alanine