冻干再水化血小板结构与功能研究

Structure and function study of rehydrated-lyophilized platelets

目的探索血小板冻干保存及再水化后结构与功能的改变。方法选择过期的冻干再水化血小板为实验对象,以新鲜血小板作为对照,分别用血小板计数仪、扫描电镜、透射电镜、血小板聚集仪和三色流式细胞仪检测血小板的回收率、稳定性、形态、超微结构、聚集反应和膜表面糖蛋白的表达。结果实验组血小板计数明显下降,血小板回收率为49.36%,且随着血小板放置时间的延长,血小板回收率逐渐下降,血小板透光性较差,分泌颗粒数目较少。实验组凝血酶、ADP、胶原及瑞斯托霉素的最大聚集率均与对照组有明显统计学差异(P<0.05)。凝血酶作用后CD62p再表达率为44.02%,显著低于对照组70.31%(P<0.05),PAC-1再表达率为38.90%,显著低于对照组65.28%(P<0.05)。结论过期血小板冻干再水化后的恢复率较低,稳定性较差,结构和功能均低于新鲜血小板,但冻干再水化血小板仍具有一定的活性,为新鲜血小板的冻干保存提供了基础。

Objective To explore the structural and functional changes of rehydrated-lyophilized platelets. Methods The rehydrated-lyophilized out-of-date platelets were chosen as experimental group and fresh platelets as control group. The recovery rate, the stability, morphology, ultrastructure, aggregation reaction and the expression of membrane surface glycoprotein of platelets were detected by the platelet counter, scanning electron microscopy, transmission electron microscopy, platelet fluorescent aggregometer and three color flow cytometry instrument respectively. Results The platelet count was significantly decreased in experimental group, platelet recovery was 49.36% which decreased gradually with the extended platelet placing time. The maximum aggregation rates of thrombin, ADP, collagen and ristomycin had significantly statistical difference between two groups（P〈0.05）.CD62 p and PAC-1 expression rate were 44.02% and 38.90%, respectively after the action of thrombin, the CD62 p and PAC-1expression rate were 70.31% and 65.28%, respectively in control group. Conclusion The recovery rate of rehydrated-lyophilized out-of-date platelets is low. The stability is poor. The structure and function are lower than those of fresh platelets, but it still has a certain activity and can provide a basis for freeze-drying preservation of fresh platelet.