| Objective:1.Investigate the effects of FS-AMB on platelet function in rats with coronary heart disease.2.Identify the differential platelet proteins in patients with coronary heart disease with cold phlegm blockade syndrome before and after the intervention of FS-AMB.Analyze the effect of differential proteins on platelet activation to find new potential targets of antiplatelet drugs,and to provide experimental basis for the clinical application of FS-AMB in antiplatelet therapy.Methods:1.The rat model of coronary heart disease was established feeding with high-fat diet and injecting pituitrin,and the 30 rats were randomly divided into experimental groups and control group,treated with low,medium and high doses of FS-AMB,aspirin and normal saline in vivo and in vitro.Extract the platelet rich plasma and detective the effects of the above drugs on platelet aggregation rate and coagulation function.2.Collect and extract the platelet of 24 patients with coronary heart disease with cold phlegm blockade syndrome,which were divided into three groups: Control group、FS-AMB group and Aspirin group.Intervene the Group X and Group A with optimal dose of FS-AMB and aspirin respectively.Two dimensional gel electrophoresis and proteomic technology were combined to identify the differential proteins among the three groups.Figure out the effects of differential proteins on the activization of platelet and analyze the mechanism of FS-AMB intervention in coronary heart disease with old phlegm blockade syndrome.Results:1.FS-AMB at different concentrations significantly inhibited ADP-induced platelet aggregation and prolonged TT,PT and APTT and reduced the Fib content in plasma.The inhibiting effect was dependent with a certain concentration.2.The platelet protein gel maps of Control group、FS-AMB group and Aspirin group were obtained by two-dimensional gel electrophoresis.Comparing FS-AMB group with Control group,21 different spots were obtained,which included three triple different spots.After mass spectrometry identification and database search,the triple differential proteins were identified as actin-cytoplasmic 1,myosin-alpha-3 chain and keratin II-cytoskeletal 1.Comparing Aspirin group with Control group,40 different spots were obtained,which included eleven triple different spots.After mass spectrometry identification and database search,the top five triple differential proteins were identified as apolipoprotein A1,actin,keratin Ⅰ-cytoskeletal 10,keratin Ⅱ-cytoskeletal 1 and keratin Ⅰ-cytoskeletal 10.Comparing FS-AMB group with Aspirin group,27 different spots were obtained,which included eleven triple different spots.After mass spectrometry identification and database search,the triple differential proteins were identified as keratin Ⅱ-cytoskeletal 1,keratin II-cytoskeletal 2,keratin Ⅰ-cytoskeletal 10,apolipoprotein A1 and keratin II–cytoskeletal 1.Conclusion:1.FS-AMB can inhibit ADP-induced platelet aggregation and coagulation function of rats with coronary heart disease,and has the function of promoting blood circulation and removing blood stasis.This result is a supplement to the traditional effect of FS-AMB and broadens the way of clinical medication of FS-AMB.2.FS-AMB could inhibit the activation of platelet by decreasing the expression of actin and myosin in platelets,so as to achieve the purpose of intervening the coronary heart disease with cold phlegm block syndrome,and its intervention mechanism is different from that of western medicine aspirin.This provides an experimental basis for the clinical application of FS-AMB in antiplatelet therapy of coronary heart disease. |
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