冻干血小板保护液优化的实验研究  

Protective liquid optimization for platelet lyophilization

作　　者：魏淑贞 施琳颖[2] 周谋[2] 林放[2] 吴红宜 李蕾 李荣娟 单桂秋[1,2] WEI Shuzhen;SHI Linying;ZHOU Mou;LIN Fang;WU Hongyi;LI Lei;LI Rongjuan;SHAN Guiqiu(Guangzhou University of Chinese Medcine , Guangzhou 510403, China;Department of Blood Transfusion, Guangzhou General Hospital of Guangzhou Military Command.)

机构地区：[1]广州中医药大学,广东广州510403 [2]广州军区广州总医院输血科

出　　处：《中国输血杂志》2018年第4期350-355,共6页Chinese Journal of Blood Transfusion

基　　金：军队后勤科研计划项目(BGZ15C002);广州市科技计划项目《(201508020115)

摘　　要：目的优化冻干血小板保护液,减少冻干过程对血小板的损害,有效保护其功能。方法采用3因素3水平的正交设计法,对血小板膜保护剂、血小板激活抑制剂、蛋白质类保护剂的3类成分组成的保护液进行优选。检测血小板最大聚集率、血小板活化标志PAC-1、CD62P的阳性表达率和血小板回收率,通过比较其差异,筛选出本实验的最优冻干血小板保护液。以优化保护液对血小板处理后进行冻干,并以此为实验组,与新鲜血小板和原保护液处理后冻干血小板进行比较验证,验证指标与优选指标相同,比较3者4项指标之间的差异。结果正交设计试验结果显示：血小板最大聚集率组间最高（74.33±24.01）%,血小板活化标志PAC-1和CD62P低于9组平均水平,血小板回收率高于9组平均水平。以主要体现血小板功能的血小板最大聚集率为优先指标,结合其余3项检测指标进行综合分析,在原保护液的基础上,筛选出添加了2%DMSO、第2信使调节剂、2 mmol/L维生素B6,75%的PPP保护液组为最优组合。验证实验结果显示,优化保护液组血小板最大聚集率（76.12±6.02）%,与原保护液组相比,差异有统计学意义（P〈0.05）,与新鲜血小板组相比,差异没有统计学意义（P〉0.05）。最优保护液组血小板激活标志PAC-1和CD62P阳性表达率分别为（3.23±0.49）%和（36.83±8.21）%,2者与原保护液组、新鲜血小板组相比,差异无统计学意义（P〉0.05）;最优保护液组冻干血小板复水化后血小板回收率为（85.90±2.24）%,与原保护液组相比,差异无统计学意义（P〉0.05）。结论本实验优化的血小板保护液优于原保护液,明显提高了冻干血小板复水化后最大聚集率,能更好地保护血小板的功能,但对血小板冻干过程的保存损害问题没有得到更好的改善,有待下一步深入研究。Objective To perform protective fluid optimization for platelet lyophilization, aiming to reduce the damage caused to platelets and to effectively maintain their biological functions. Methods An orthogonal design of 3 variables and 3 levels are used to optimize the solution formula： The platelet membrane protector,the platelet activation inhibitor and the protein protectant. The optimized combination was acquired by comparing the differences of the maximum platelet aggregation rate,the positive expression rate of platelet activation markers（PAC-1,CD62 P） and the recovery rate of platelets. The experimental group with the optimized solution treatment was compared with the fresh platelet group and the original protective solution group. The verification indexes were the same and the differences of the four indexes are compared among the 3 groups. Results The optimization result of the orthogonal design showed that the highest platelet aggregation rate was at（74. 33＋ 24. 01）% with lower-than-average platelet activation markers expressed and a higher-than-average platelet recovery rate. The maximum aggregation rate of platelets,which mainly defines platelet functions,was weighed as the priority index.The optimized combination acquired was： 2% DMSO,second messenger regulator,2 mmol/L vitamin B6 and 75% PPP. For the experiment group,the maximum platelet aggregation rate was the highest among the 3 at（76. 12±6. 02） % with statistical significance compared to the original protective solution group（P〈0. 05） and,remarkably,no statistically significant differences compared to the fresh platelet group（P〉0. 05）. The platelet activation marker expression status presented no statistical significance among the 3 groups. The rate of platelet recovery was（85. 90＋ 2. 24） % in the experiment group with no statistically significant differences compared to the original protection solution group（P〉0. 05）. Conclusion The optimized platelet protective solution is superior to the original protect

关 键 词：冻干血小板 血小板保护液 血小板储存损伤 正交设计法 血小板最大聚集率 

分 类 号：R331.143[医药卫生—人体生理学] R457.1[医药卫生—基础医学]