抄録
In using high pressure liquid chromatographic technique nucleosides, cyclic 3', 5'-AMP, cyclic 3', 5'-GMP and 5' mono-, 5' di-, 51trinucleotides could be analyzed. All nucleotides could be quantitated in a single step separation within 45min employing 1m mole/l potassium phosphate (pH 3.0) between 0.4mole/l potassium phosphate (pH 3.1) gradient. Half ml of plateletrich plasma with 1/10 volume of ACD or 3.8 % sodium citrate furnished an adequate sample for analysis.
Nucleotide levels in platelets from 12 healthy donors expressed in 1011 platelets were as follows: AMP 0.56 (0.22) mean (SD), ADP 2.6 (0.6), ATP 4.0 (0.5), GMP 0.32 (0.22), GDP 0.48 (0.2), GTP 0.32 (0.08), UDP 0.28 (0.04) and CTP 0.11 (0.08) micromoles.
Upon platelet aggregation without release of ADP, ATP and platelet factor 4 (primary aggregation) induced by lower concentration of ADP and epinephrine, no remarkable change of nucleotide profiles was observed. Amount of ATP, ADP, GTP and GDP in platelets was decreased down to 50% of initial values during aggregation with release reaction (secondary aggregation) by collagen, arachidonate and higher concentration of ADP and epinephrine. Ratio of ATP/ADP in platelets showed no significant change before and during release reaction. As amount of released ADP was usually higher than ATP released, these nucleotides would decrease not only by the extrusion from the platelets in the release but also by consumption during release reaction.
It is concluded that release reaction of platelets always accompanies the decrease of nucleotide levels and platelet aggregation itself does not necessarily accompany the change of nucleotide contents. Loss of nucleotides during release reaction is derived from the extrusion from the platelets and consumption in the platelets.
