Cold storage of rabbit thoracic aorta in University of Wisconsin solution attenuates P2Y(2) purine receptors.
Post-transplantation thrombosis may occur in donor segments of iliac arteries and livers following surgical removal and storage in University of Wisconsin (UW) solution for transplantation. We have previously suggested that purine receptors are vulnerable to denaturation after UW storage. The aims of the present study were to determine what particular subtypes of purine P2Y receptors in rabbit thoracic aorta deteriorate after 8 days of UW storage by studying vascular reactivity to acetylcholine, ATP, 2MeSATP and UTP. Ring segments of aortae from male New Zealand White rabbits were mounted upon fine-wire myographs and vasodilatation to the above agents tested on fresh tissue, and after 8 days of UW storage. Vasodilatation to ATP was attenuated by 100 microM L-NAME in fresh tissue suggesting that the relaxant response was, in part, due to nitric oxide (NO). P2Y-mediated relaxation to ATP was significantly attenuated by UW storage and cholinergic responses were not. This attenuated relaxation to ATP was not further attenuated by L-NAME, suggesting a loss of the NO-dependent mechanism. De-endothelialisation indicated that UTP-mediated vasorelaxation, via P2Y(2) receptors, was endothelium-dependent. Any residual endothelium-independent relaxation to UTP was abolished by UW storage and endothelium-dependent UTP relaxation was reduced to the same level as that seen in fresh, de-endothelialised tissue. In contrast responses to 2MeSATP, via P2Y(1) receptors, were predominantly endothelium-independent and were only partially attenuated by UW storage. Responses to pyridoxalphosphate-6-azophenyl-2('),4(')-disulphonic acid (PPADS) and L-NAME suggested that vasorelaxation to 2MeSATP and UTP was mediated by P2Y(1) and P2Y(2) receptors, respectively. It is therefore concluded that UW storage predominantly decreases P2Y(2) receptor-mediated vascular reactivity.
Payne
Sarah J
SJ
Liver Sciences Unit, Academic Department of Surgery, GKT School of Medicine, St. Thomas' Hospital, Lambeth Palace Road, London SE1 7EH, UK.
Benjamin
Irving S
IS
Alexander
Barry
B
D000109:Acetylcholine / Q000494:pharmacology; D000241:Adenosine; D000255:Adenosine Triphosphate / Q000031:analogs & derivatives* / Q000494:pharmacology; D000493:Allopurinol; D000818:Animals; D001013:Aorta, Thoracic / Q000187:drug effects / Q000378:metabolism* / Q000502:physiology; D003080:Cold Temperature; D005978:Glutathione; D066298:In Vitro Techniques; D007328:Insulin; D008297:Male; D019331:NG-Nitroarginine Methyl Ester / Q000494:pharmacology; D019188:Organ Preservation Solutions; D011817:Rabbits; D011887:Raffinose; D018048:Receptors, Purinergic P2 / Q000378:metabolism*; D058473:Receptors, Purinergic P2Y2; D013873:Thionucleotides / Q000494:pharmacology; D014021:Tissue Preservation / Q000379:methods*; D014544:Uridine Triphosphate / Q000494:pharmacology; D014664:Vasodilation / Q000187:drug effects
D007328:Insulin; D019188:Organ Preservation Solutions; D018048:Receptors, Purinergic P2; D058473:Receptors, Purinergic P2Y2; D013873:Thionucleotides; C055206:University of Wisconsin-lactobionate solution; D000493:Allopurinol; D000255:Adenosine Triphosphate; D005978:Glutathione; D000241:Adenosine; D011887:Raffinose; D000109:Acetylcholine; D014544:Uridine Triphosphate; D019331:NG-Nitroarginine Methyl Ester; C008501:2-methylthio-ATP