Increased hypoxic stress decreases AMP hydrolysis in rabbit heart.
OBJECTIVE
AMP conversion to adenosine by cytosolic 5'nucleotidase (5NT) or to IMP by AMP deaminase determines the degree of nucleotide degradation, and thus ATP resynthesis, during reoxygenation. To elucidate the regulation of AMP hydrolysis during ischemia, data from 31P NMR spectroscopy and biochemical analyses were integrated via a mathematical model. Since 5NT is downregulated during severe underperfusion (5% flow), we tested 5NT regulation during less severe underperfusion (10% flow) and then made the perfusate hypoxic to see if the greater stress reactivated 5NT.


METHODS
31P NMR spectra and coronary venous effluents were obtained from Langendorff-perfused rabbit hearts subjected to two 30-min periods of underperfusion (10% flow); the second period with or without additional hypoxia (30% O2). Data were analyzed with a mathematical model describing the kinetics of myocardial energetics and metabolism.


RESULTS
A single 30-min period of 10% flow causes downregulation of AMP hydrolysis and the data from the second period of underperfusion are best described by lower 5NT activity, even in the presence of extra hypoxia. Thirty percent less purines appear in the venous effluent than predicted by the phosphoenergetics (PCr and ATP) when IMP is not allowed to accumulate by the model, however the model indicates that a constant accumulation of IMP via AMP deaminase could explain the discrepancy between expected and measured purines in the venous effluent.


CONCLUSIONS
While AMP hydrolysis to adenosine is prominent in early ischemia and acts to preserve cellular energy potential, during a second ischemic period, nucleotides are conserved by the stable inhibition of AMP hydrolysis. Furthermore, during 10% flow conditions, nucleotides are conserved, possibly via an IMP-accumulatory pathway.
Gustafson
L A
LA
Center for Bioengineering, University of Washington, Seattle 98195, USA. lorig@physiol.med.vu.nl
Zuurbier
C J
CJ
Bassett
J E
JE
Barends
J P
JP
van Beek
J H
JH
Bassingthwaighte
J B
JB
Kroll
K
K
D000241:Adenosine / Q000378:metabolism*; D000249:Adenosine Monophosphate / Q000378:metabolism*; D000704:Analysis of Variance; D000818:Animals; D003198:Computer Simulation; D006868:Hydrolysis; D009682:Magnetic Resonance Spectroscopy; D008955:Models, Cardiovascular; D017202:Myocardial Ischemia / Q000378:metabolism*; D009206:Myocardium / Q000378:metabolism*; D010477:Perfusion; D011687:Purines / Q000378:metabolism; D011817:Rabbits; D013997:Time Factors
D011687:Purines; D000249:Adenosine Monophosphate; D000241:Adenosine