Bozzo J, Hernández M R, Ordinas A
Servei d'Hemoteràpia i Hemostàsia, Hospital Clínic i Provincial, Villarroel, Barcelona.
Sangre (Barc). 1996 Feb;41(1):13-8.
The modulating role played by red cells on platelet function is well known, yet its mechanism of action in the presence of antiaggregating agents is not clearly defined. We tried to assess the influence of aspirin (ASA) or dipyridamole (DIP) treated red cells on platelet aggregation in plasma, with the samples in static conditions and under flow in a perfusion system.
Platelet-rich plasma (PRP) was adjusted to 4 x 10(9)/L. Red cells were washed with a 0.9% sodium chloride and 5mM glucose solution, then resuspended in PRP until a 40% haematocrit was attained. The drugs were added separately or together and the mixture was incubated for 10 minutes at 37 degrees C. Three series of tests were carried out: a) with reconstituted blood; b) with red cells alone; c) with the red cells washed in a 10-fold volume so as to eliminate all the drugs. The samples were divided in two parts: one was let to rest and the other was circulated through a perfusion system with a peristaltic pump. Simultaneous tests with untreated red cells and PRP were run as control in each session. A multichannel aggregameter was used to assess platelet aggregation according to Born's turbidimetric method. Variance analysis (ANOVA) or Student's t test were used for the statistical evaluation.
Platelets attained from control samples under flow conditions showed partial inhibition of the maximal aggregation (58.02 +/- 9.92%). The inhibitory power of ASA was higher than that of DIP in all cases. Aggregation was totally inhibited by ASA from the beginning in treated whole blood, but the inhibitory effect appeared later when working with red cells alone, suggesting that red cells must retain part of the drug, which cannot exert its effect until it has been released after several minutes. The inhibition of platelet aggregation when working with washed treated red cells was more effective under flow conditions, chiefly with combined treatment with ASA+DIP.
These studies stress the importance of flow conditions in the red cells-platelets interaction and in the regulation of the therapeutic agents used to modify platelet function.
红细胞对血小板功能的调节作用已为人熟知,但其在抗聚集剂存在时的作用机制尚不清楚。我们试图评估经阿司匹林(ASA)或双嘧达莫(DIP)处理的红细胞对血浆中血小板聚集的影响,样本分别处于静态条件和灌注系统中的流动状态。
将富血小板血浆(PRP)调整至4×10⁹/L。红细胞用0.9%氯化钠和5mM葡萄糖溶液洗涤,然后重悬于PRP中,直至血细胞比容达到40%。分别或联合加入药物,混合物在37℃孵育10分钟。进行了三组试验:a)用重构血液;b)仅用红细胞;c)用10倍体积洗涤的红细胞以去除所有药物。样本分为两部分:一部分静置,另一部分通过蠕动泵在灌注系统中循环。每次试验均同时进行未处理红细胞和PRP的对照试验。使用多通道聚集仪根据博恩比浊法评估血小板聚集。采用方差分析(ANOVA)或学生t检验进行统计学评估。
在流动条件下,对照样本中的血小板显示最大聚集受到部分抑制(58.02±9.92%)。在所有情况下,ASA的抑制能力均高于DIP。在处理的全血中,ASA从一开始就完全抑制聚集,但单独使用红细胞时,抑制作用出现较晚,这表明红细胞必须保留部分药物,药物在数分钟后释放才能发挥作用。在用洗涤处理的红细胞进行试验时,在流动条件下对血小板聚集的抑制更有效,主要是联合使用ASA+DIP时。
这些研究强调了流动条件在红细胞 - 血小板相互作用以及调节用于改变血小板功能的治疗药物方面的重要性。