Vecino A M, Navarro J L, Cesar J M
Departamento de Hematología, Hospital Ramón y Cajal, Madrid, Spain.
Rev Esp Fisiol. 1996 Mar;52(1):45-8.
Isolated platelets from samples with low counts produce technical problems. Albumin gradient (AG) has been shown to be useful for this purpose, preserving the aggregating response of these cells. The influence of this method in the enzymatic pathways that regulate the platelet activation is studied. Platelets were isolated by either AG or conventional centrifugation methods and labelled with C-14-arachidonic acid (C-14-AA). Isolated platelets were activated with thrombin (5 U/ml) and lipids were extracted according to Bligh and Dyer. Platelet phospholipids and prostanoids were resolved by TLC. The incorporation of C-14-AA by platelets was similar by both methods (31.7 +/- 18% versus 47.2 +/- 6.9%), as well as the distribution of C-14-AA in the five major platelet phospholipids. Formation of radioactive thromboxane B2, hydroxyheptadecatrienoic acid and hydroxyeicosatetraenoic acid by activated platelets was also similar by both methods. These findings suggest that platelet isolation by albumin gradient preserves the enzymatic pathways responsible for the activation of these cells.
从血小板计数低的样本中分离出的血小板会产生技术问题。白蛋白梯度（AG）已被证明在此方面有用，可保留这些细胞的聚集反应。本研究了该方法对调节血小板活化的酶促途径的影响。通过AG或传统离心方法分离血小板，并用C-14-花生四烯酸（C-14-AA）标记。分离出的血小板用凝血酶（5 U/ml）激活，并根据布莱和戴尔法提取脂质。血小板磷脂和前列腺素通过薄层层析法分离。两种方法使血小板对C-14-AA的摄取相似（分别为31.7±18%和47.2±6.9%），C-14-AA在五种主要血小板磷脂中的分布也相似。两种方法激活的血小板形成放射性血栓素B2、羟基十七碳三烯酸和羟基二十碳四烯酸的情况也相似。这些发现表明，通过白蛋白梯度分离血小板可保留负责这些细胞活化的酶促途径。
