储存损伤对血小板超微结构、磷脂及聚集功能的影响

The Changes of Lipids, Ultrastructure and Aggregation in Platelet Storage Lesion

目的对保存期间血小板脂质、超微结构及功能的改变进行观察分析，探讨储存损伤对血小板的影响。方法２０名健康献血者，常规采血，经二次离心获得浓缩血小板，将每单位浓缩血小板分装成两份，分别编为甲、乙两组，每组２０个样本。甲组于２２℃水平振摇保存，分别在３天、５天时取样检测；乙组采用ＤＭＳＯ作为防冻剂，终浓度为５％，－８０℃保存，分别在１月、３月、６月时取样测定；采血当天的新鲜血小板为对照组。采用高效液相色谱法（ＨＰＬＣ－ＵＶ）分析血小板磷脂的变化，电镜观察血小板超微结构的改变。结果正常人新鲜血小板中四种主要磷脂：磷脂酰胆碱（ＰＣ），磷脂酰乙醇胺（ＰＥ），磷脂酰丝氨酸（ＰＳ），磷脂酰肌醇（ＰＩ）的含量依次为ＰＣ＞ＰＥ＞ＰＳ＞ＰＩ，各占总磷脂含量的３７％，２７％，１２％，７％左右。血小板在２２℃水平振摇保存３天、５天，磷脂丢失率为５．８％，３１．８％；－８０℃冷冻保存（防冻剂为ＤＭＳＯ）１～６月，磷脂丢失率约为３３～３６％左右，且不随冷冻时间延长而增加。透射电镜观察血小板超微结构，发现血小板出现激活信号，如由盘状向球形转变，伪足延伸，开放小管系统（ＯＣＳ）肿胀，储存颗粒丧失，空地形成等。血小板聚集实验显示：血小板对聚集?

objective: The purpose of this project was to study the storage lesion of platelet and to investigate the changes of platelet lipids, ultra-structure and aggregation. Methods: 20 units of platelet were divided into two groups. One was stored at 22℃ and esamined at 3,5 days poststorage. Another added DMSO (5% terminal concentration)was stored at-80℃ and examined at 1, 3 and 6 months poststorage. The fresh platelet was as control. The platelet lipids were assayed by HPLC. The platelet ultra structure was investigated by electron microscope. Results: The loss rates of platelet lipids at 22℃ were 5.8%-31.8% after 3, 5 days respectively. The loss rates of lipids at -80℃ were 33%-36% after 6 months storage and did not change anymore there after. The ultra-structure alterations were occurred at 22℃ and - 80℃ groups in storage. The morphological alterations were expressed as signs of activation. The piatelet aggregative test showed the decrease of reaction to agonist ADP. This result showed the functional alteration of platelet. Conclusion: The functional alterations of platelet are due to the lipids loss and morphologi cal alterations in platelet preparation and storage.