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Citation Information: J Clin Invest. 1985;76(1):233-240. https://doi.org/10.1172/JCI111952.
Abstract
Methods were developed for measuring changes in platelet sensitivity to a release-inducing stimulus and in platelet cyclic AMP in fresh whole blood samples from rabbits. These techniques permitted detection of the effects of exogenous and endogenous prostacyclin on circulating platelets. In these methods, rabbit platelets were labeled in vitro by incubation with [14C]serotonin and [3H]adenine and then transfused into other rabbits. Release of platelet [14C]serotonin by a standard dose of synthetic platelet-activating factor (40 pmol/ml) and the platelet cyclic [3H]AMP levels were then measured in citrated blood from the conscious animals within 2 min of arterial puncture. Bolus intravenous injections of prostacyclin (1-10 nmol/kg) caused concentration-dependent increases in platelet cyclic AMP after 2 min, which decreased approximately 75% by 5 min, and disappeared after 30 min. Significant inhibition of the platelet release reaction was detected 2 min but not 5 min after injection of 10 nmol of prostacyclin per kilogram. With lower doses, significant enhancement of the release of [14C]serotonin was observed after 5 min. Similar changes in platelet responsiveness and cyclic [3H]AMP were observed after release of endogenous prostacyclin by intravenous injection of angiotensin II (5 nmol/kg); inhibition of the release of [14C]serotonin after 2 min was followed by potentiation after 5 min, though platelet cyclic [3H]AMP remained above control values. In these experiments, the time course of the changes in platelet cyclic [3H]AMP correlated closely with values for blood prostacyclin obtained previously (Haslam, R.J., and M.D. McClenaghan, 1981, Nature [Lond.]., 292:364-366). Prostacyclin also had a biphasic effect on the release of [14C]serotonin when added to citrated blood in vitro, though both the increase in sensitivity to platelet-activating factor and the return of platelet cyclic [3H]AMP towards control values took place more slowly. At all times, addition of platelet-activating factor decreased platelet cyclic [3H]AMP towards but not below the control level observed in the absence of prostacyclin. Our results indicate that although transient increases in platelet cyclic AMP cause an immediate decrease in platelet responsiveness in vivo or in vitro, a period of enhanced platelet sensitivity follows as platelet cyclic AMP falls.
Authors
M Vanderwel, R J Haslam
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