Full Text
Abstract
Introduction. Platelet activation stimulates their aggregation and secretion associated exocytosis of intracellular granules. Ag- gregation is usually studied by turbidimetric method and granule exocytosis by detecting expression of their markers on plate- let surface using flow cytofluorimetry.
Aim: сomparison of the sensitivity of two methods for evaluation of platelet activation, studies of their aggregation and expres- sion of intracellular granule markers.
Materials and methods. Healthy donors’ platelets were activated by ADP and thrombin receptor activating peptide (TRAP). Plate- let aggregation was investigated in platelet rich plasma by turbidimetric method, assessed by the maximal level of light trans- mission (T% max). Expression of granule markers was detected in whole blood by flow cytofluorimetry registering the percent of platelets stained with antibodies against the markers of alpha-granules (CD62P+) and dense granules (CD63+).
Results. In all donors 20 μM ADP and 10 μM TRAP stimulated strong irreversible platelet aggregation (62.1 ± 10.3 and 65.1 ± 5.1 T% max). Mean aggregation levels were lower when it was stimulated by 2.5 μM ADP (30.9±23.2 T% max) and 1 μM TRAP (24.4±29.1 T% max). Expression of granule markers was maximal at platelet activation by 10 μM TRAP (77.6 ± 13.7% CD62P+ and 73.9 ± 13.3% CD63+), lower at activation by 1 μM TRAP (46.3 ± 25.1% CD62P+ and 38.3 ± 23.8% CD63+), more lower at activation by 20 μM ADP (19.8 ± 8.5% CD62P+ and 13.6±5.4% CD63+), and minimal at activation by 2.5 μM ADP (8.0±4.2% CD62P+ and 8.3±3.3% CD63+). Epinephrine (20 μM) did not stimulate expression of granule markers but at combined addition with 20 μM ADP increased its level by 1.9 for both mark- ers, CD62P and CD63.
Conclusion. Platelet aggregation is more sensitive method than detection of expression of granule markers when platelets are ac- tivated by ADP. Both methods demonstrate about the same sensitivity when platelets are activated by TRAP. Expression of granule markers is increased at combined addition of ADP and epinephrine although epinephrine by itself fails to stimulate their expression.
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