【摘要】 目的建立流式细胞仪测定血小板胞浆游离钙离子浓度[Ca2+]i的方法。方法用Fluo 3-AM作为钙指示剂,在有或无细胞外钙[Ca2+]o存在的条件下,测定静息和凝血酶激活状态下人血小板胞浆游离钙离子浓度[Ca2+]i的变化。结果Fluo 3-AM标记的血小板的平均荧光强度明显增加。凝血酶使负载Fluo 3-AM标记的血小板胞浆[Ca2+]i明显增加,且显示剂量依赖和对[Ca2+]o的依赖。结论凝血酶引起血小板胞浆[Ca2+]i增加的来源主要是[Ca2+]o,可能也有部分是内钙释放的参与。更多还原

【Abstract】 Objective To set up a method to determine cytoplasmic free calcium [Ca²⁺]_i in human platelets with flow cytometry.Methods As a calcium indicator,Fluo 3-AM was used to determine the changes,induced by thrombin,of the level of platelet plasma free calcium [Ca²⁺]_i in presence or absence of outside calcium [Ca²⁺]_o,in order to elucidate where the platelet plasma free calcium [Ca²⁺]_i mainly come from and it’s role in the activity of platelets.Results The geometric mean of fluorescence intensity of the platelet,labeled with Fluo 3-AM,was increased obviously.The concentration of platelet plasma free calcium [Ca²⁺]_i was increased significantly by thrombin in a dose-and [Ca²⁺]_o-dependent manner.Conclusion The increased level of platelet plasma free calcium [Ca²⁺]_o,induced by thrombin,mainly comes from [Ca²⁺]_o and partly from the released [Ca²⁺]_o of intra-platelet probably.更多还原