摘要
目的建立单细胞血小板钙浓度测定方法,探讨正常人服抗血小板功能药物前后,血小板钙离子浓度变化及相应功能变化的关系。方法利用特异荧光染料 Flou-3对血小板内钙离子染色,并在激光共焦显微镜下,观察同一个血小板在激活前后钙离子浓度、形态及聚集率变化。结果加入二磷酸腺苷(ADP)激活后血小板钙浓度是激活前的269.3%。激活前,血小板形态呈圆形或椭圆形,边缘较光滑,无空泡。加入 ADP 后,形态不规则,可见手镜样突起,突出部分为大空泡。服药第二天,ADP 激活后的血小板钙浓度仅为激活前的76.3%,血小板形态无变化,聚集率明显减低。结论正常人血小板经 ADP 激活后,胞浆内钙离子浓度明显升高,并引起相应的形态变化和功能的改变,阿司匹林可抑制这种变化。应用激光共焦显微术探讨单个血小板钙浓度变化,证实其可以直观血小板钙浓度变化,并能同时观察这种变化与血小板形态和功能的关系,可以作为研究血小板功能及抗血小板药物疗效的一种新的实验手段。
Objective To observe the relation between Ca^(2+) concentration in a single platelet and platelets aggregation in the normal before and after taking aspirin.Methods Ca^(2+) concentration in a single Flou-3-1abeled platelet was observed with laser scanning confocal microscope(LSCM).The platelet shape change and aggregation were observed simultaneously.Results Without aspirin,Ca^(2+) concentration in a single platelet rose sharply and the shape of the platelet changed distinctly after ADP stimulation.With aspirin,the platelet had indistinetly rising Ca^(2+) concentration,no shape change and reduced aggregation rate after ADP stimulation.Conclusions 1.The normal platelet had remarkably rising Ca^(2+) concentration,and distinct shape and aggregation change after ADP stimulation.Aspirin in- hibited the increase of Ca^(2+) concentration in the platelet and the aggregation of platelets after ADP stim- ulation.2.We observed Ca^(2+) concentration in a single platelet with ISCM for the first time in China, and this method is a new way to study platelet function,pathogenic factors and therapeutic drugs.
