冷冻干燥保存血小板实验探究.doc

冷冻干燥保存血小板实验探究 　 作者：刘景汉，周俊，王冬梅，欧阳锡林，邢颜超，卢发强 【摘要】 本研究探讨冷冻干燥保存血小板的方法，以获得可长期冻干保存的血小板制品，使之能在常温条件下保存、占用空间小、重量轻、便于长距离运输，能够满足突发事件和战伤救治的需要。在冷冻干燥保存过程中添加血小板可逆性激活抑制剂、DMSO和海藻糖等低温保护剂，进行预处理、冷冻、一级干燥、二级干燥，再水化，并同时测定血小板回收率，凝血酶聚集反应，促凝血功能，CD62p表达率和PAC1表达率等。结果表明：血小板回收率为56.29％，其对凝血酶的聚集反应与对照组无明显差异，对ADP和丙基没食子酸诱导的聚集反应较对照组分别降低49.34％和26.25％，促凝血功能与对照组比较也无统计学差异；冻干血小板CD62p表达率为42.36％，PAC1表达率为2.12％，凝血酶激活后CD62p再表达率为50.88％，PAC1再表达率为54.55％。结论：添加血小板可逆性激活抑制剂，海藻糖和DMSO后的冻干血小板，其聚集活性和促凝血功能与新鲜血小板无明显差异，血小板可逆性激活抑制剂降低了冻干血小板的CD62p表达，增强了冻干血小板的生存能力，因而延长了其生存时间，因此可以该冻干方法为基础进一步提高冻干保存血小板的效率。 【关键词】 可逆性激活抑制剂 Experimental Study on Lyophilization of Platelets Abstract The aim of this study was to search a procedure of platelet lyophilization and find a way of longterm sto rage of human platelets at normal temperature with smaller size and lighter weight, to be convenient to transport at long distance thus to meet the demands in accidents and war time. Human platelets were pretreated by freezing, the first and the second desiccation, and were added with reversible activationinhibitors of platelets, DMSO and trehalose, then were rehydrated. At the same time, the recovery rate of platelets, platelet maximal aggregation induced by thrombin, coagulation of platelets, CD62p expression and PAC1 expression were assayed. The results indicated that the recovery rate of the plalelets was 56.29%. The platelet maximal aggregation induced by thrombin had no significant difference between lyophilized platelets and the fresh plateletrich plasma (FPRP), but the aggregation of platelets induced by ADP or propyl gallate was decreased by 49.34% and 26.25%. Coagulation of the lyophilized platelets was not significantly different from FPRP. CD62p expression of the lyophilized platelets (42.36％) was higher than that in FPRP while PAC1 expression was 2.12%. CD62p reexpression rate induced by thrombin was 50.88% and PAC1 reexpression was 54.55%. It is concluded that the ability of recovered lyophilized platelets added with reversib