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凝血酶对荧光标记的人单个血小板游离[Ca~(2+)]和膜电位的动态影响

庞建新, 单春文

(第一军医大学药理教研室，广州510515)

摘要

该文将fluo-3和di- BA-CQ (3)荧光标记的血小板固定于纤维蛋白原表面，以570型粘附式细胞仪(ACAS 570)动态观察了凝血酶激活的人单个血小板细胞内游离[Ca~(2+)I钙离子浓度)和膜电位的变化。静息状态时细胞游离[Cal+口和膜电位荧光较低，波动不明显。当加入0. 1 U/MI凝血酶激活时，[Ca~(2+)]}(细胞内游离钙离子浓度)与膜电位迅速升高，随后[Ca 2+]出现反复振荡，幅度达约500荧光单位，而膜电位基本上保持峰值水平。[Ca ~(2+)];升高与膜电位变化在时间和程度上不一致。本文结果提示，凝血酶引起血小板「Ca ~(2+)]振荡和膜去极化，后者不是Cal~(2 +)内流引起的。

关键词： 单个血小板; 游离Ca2+; 膜电位; 凝血酶

EFFECT OF THROMBIN ON CYTOSOLIC CALCIUM AND MEMBRANE POTENTIAL IN SINGLE HUMAN PLATELETS

PANG JIAN-XIN , SHAN CHUN-WEN

(Department可Pharnwcology, First Military Medical University, Qngzhou 510515)

Abstract

Fluo-3 and d;-BA-Cg (3)-loaded human platelets were immobilized on a fibrinogen-coated surface and the cytosolic free [Ca2+]、and the membrane potentials were measured in single thrombin-activated platelets by ACAS-570 (Adherent Cell Analysis and Sorting Cytometer ).Fluorescence of fluo-3 and d;-BA-C4 (3) was low in rest in-f7 mobilized platelets without obviously repetitive spiking. When the platelets were activated by thrombin，a rapid rise of cytosolic ECa2+I; and membrane potential was observed.Subsequently，oscillations in〔Ca2+]，to an extent of about 500 fluorescent units were
observable，whereas the membrane potential remained at the peak level，i. e，the change of[CaZ+」，in no way of accord with that membrane potential. The above results indicate that the thrombin induced membrane depolarization is not due to influx of Ca2+.

Key words: hrombin;single platelet;cytosolic free-「CaZ+;membrane potential

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引用本文：

庞建新, 单春文. 凝血酶对荧光标记的人单个血小板游离[Ca~(2+)]和膜电位的动态影响[J]. 生理学报 1996; 48 (3): .

PANG JIAN-XIN , SHAN CHUN-WEN. EFFECT OF THROMBIN ON CYTOSOLIC CALCIUM AND MEMBRANE POTENTIAL IN SINGLE HUMAN PLATELETS. Acta Physiol Sin 1996; 48 (3): (in Chinese with English abstract).