Research On The Membrane Protein Of Platelet Microparticles By Flowcytometer

Subject We try to establish the method for the membrane protein on platelet microparticles(PMP) by flow cytometry, and compare the percent of PAC-1 + PMP or CD62p+ PMP by activating platelet with different agonists, and approach the relation between the membrane protein on platelet microparticles and coronary heart disease or hypertension, and approach the dynamic change in the membrane protein on platelet microparticles of patient with anti-platelet therapy, and look for a new test which are helpful to diagnosis the coronary heart disease and supervise the anti-platelet therapy.Method We examine the percent of PAC-l+ PMP or CD62p+PMP by activating platelet with varying concentrations of ADP (5 u M, 10 u M, 20 u M) , thrombin (0.1U/ml, 0.5U/ml, 1U/ml) , collagen (5 u g/ml, 10 w g/ml, 20 n g/ml) in the same sample collected from normal health people(n=10). We examine the percent of PAC-1+ PMP or CD62p+ PMP by activating platelet with 20 u M ADP in the blood collected from the patient with coronary heart disease (n=12) and hypertension(n=10). We collected blood samples from the patient with coronary heart disease(n=9) treated with ticlopidine (250mgX2 in the first and second day,250mg/day from the third to sixth day)in different time(before treatment, and 4d, 5d, 6d, 9d, lOd after treatment), and examine the percent of PAC-1+ PMP or CD62p+ PMP in the samples by activating platelet with 20 u M ADP.Result As the concentrations of ADP and thrombin and collagen increased, the percent of PAC-l+ PMP or CD62p+PMP increased, and there is the obvious difference in the percent of PAC-l+ PMP or CD62p+PMP caused by different concentration of the same agonist, but there is not apparent difference between PAC-1 and CD62p on the same concentration of the same agonist. By activating platelet with 20 y M ADP, the percent of PAC-l+ PMP or CD62p+ PMP in coronary heart disease3group (85.1%?3.7%, 85.9%?.8%) are higher than normal group (75.5%?4.4%,76.3% 5.3%) (p<0.001),so are hypertension group (86.3% ?5.8%, 86.4% 5.7%) .But there is not apparent difference between coronary heart disease group and hypertension group. By activating platelet with 20 u M ADP, the percent of CD62p+ PMP at different time{before treatment, and 4d, 5d, 6d, 9d, lOd after treatment) are 84.3%?.6%, 81.4%?.4%, 70.3%?.7%, 70.6%?.2%, 83.5% ?.8%, 85.4%?.1%, asof PAC-1+ PMP are 85.3%?.5%, 82.5%?.2%, 72.3% ?.5%, 72.4%?.3%, 85.4?.4%, 86.0+3.8%. The percent of CD62p* PMP and PAC-r PMP in 5d obviously increase(p<0.001), and restore in 9d There is not apparent difference between PAC-1 and CD62p at the same time. Conclusion In the course of activating platelet, as the concentrations of ADP and thrombin and collagen increased, the percent of CD62p+ PMP and PAC-1+ PMP increased. There is some relation between the membrane protein on platelet microparticles and coronary heart disease or hypertension. There is the dynamic change in the membrane protein on platelet microparticles of patient with anti-platelet therapy. Examining the percent of CD62p+ PMP and PAC-rPMP is helpful to diagnosis the coronary heart disease and supervise the anti-platelet therapy.