血小板4℃冷藏保存方法的实验研究

Research the Method in 4℃ Cold-stored of Platelets

目的探讨以尿嘧啶二磷酸半乳糖为添加剂的血小板冷藏保存方法。方法采集兔心脏血,按常规方法制备浓缩血小板悬液(PCs),在悬液中添加5g/L的尿嘧啶二磷酸半乳糖(UDP-Gal),于37℃孵育2h后,放4℃冰箱储存10d。而后观察血小板(PLT)数量、血小板平均体积(MPV)、血小板体积分布宽度(PDW)、血小板聚集功能(PagF)、血小板促凝活性(APCT)和血小板膜AnnexinV结合率的变化;将添加UDP-Gal冷藏保存的兔血小板输入兔血小板减少症模型体内,检测输注后1h和24h兔耳出血时间和血小板回收率(PPR)。结果添加UDP-Gal冷藏保存10d的PCs中PLT数量、MPV、PDW、APCT与新鲜PCs相比,差异均无显著性(P>0.05);其血小板凋亡率与新鲜PCs相比有所增高,但显著低于冷藏对照组(P<0.01),血小板聚集率(诱聚剂为阳离子没食子酸丙酯,C-PG)减低,但仍能保持在新鲜血小板的50%以上;而冷藏对照PCs的PLT数量显著减少,MPV、PDW显著增大,且血小板促凝血活性减低,血小板PagF丧失。UDP-Gal冷藏保存的兔血小板可明显缩短兔耳出血时间(P<0.01),而冷藏对照PCs输注前后兔耳出血时间的变化无显著性(P>0.05);新鲜血小板组、UDP-Gal+冷藏血小板组、冷藏血小板对照组输入兔血小板减少症模型体内1h和24h的PPR分别是66.1%±0.5%、47.8%±0.6%,60.9%±0.3%、41.6%±0.4%、47.7%±0.5%、9.4%±0.5%,UDP-Gal+冷藏兔血小板组与新鲜兔血小板相比,其PPR的差异无显著性(P>0.05),UDP-Gal+冷藏兔血小板组和新鲜兔血小板组的PPR均显著高于冷藏血小板组(P<0.01)。结论添加尿嘧啶二磷酸半乳糖的兔血小板冷藏保存后的体内外功能良好。

Objective To study the method for storing platelets in cold-stored by Adding Uridine Diphosphate Galaetose. Methods After treatment with 5 g/L uridine diphosphate galactose（UDP-Gal/ at 37℃ for 2 hours,the rabbit platelet concentrates （PCs, 1.5 × 10^12/L ）were stored in 4℃refrigerator for up to ten days,the biochemistry and function in vitro （including platelet count, mean platelet volume, platelet distributing width, platelet aggregation function, platelet procoagulation activity,platelet hemostatic efficacy was evaluated after transfused into model of rabbit thrombocytopenia. Results There was not significant difference for PLT count,MPV, PDW and APCT between UDP-Gal-treated chilled platelet group and fresh plat.elet group （P〉0.05）. On the contrary,platelet count, decreased significantly. MPV,PDW rised and APCT went down in chilled platelet group compared with fresh platelet group （P〈0.01）. Platelet apoptosis rate increased in UDP-Gal-treated chilled group compared with fresh platelet group（P〈0. 051, but was significantly lower than that in chilled platelet group （P〈0. 011. Although PagT （inducer, cationic propyl gallate ） decreased in UDP-Gal-treated chilled group, it was still above 50% of fresh platelet group. The rabbit ear bleeding time was significantly shortened after transfusion of UDP-Gal-treated chilled platelets （P〈0. 011. On the contrary,it had less change in chilled platelets（P〉 0.05）. PPR were 66. 1%±0. 5% ,47.8%±0.6% ）60.9%±0. 3% ,41.6%±0. 4% ;47. 7%±0. 5% ,9.4%±0.5% respectively in fresh platelet group, UDP-Gal-treated chilled platelet group and chilled platelet group,it had no statistical difference between UDP-Gal-treated chilled platelet group and fresh ptatelet group （P〉0.05）,and those in both groups were much higher than that in chilled platelet group （P〈0. 01）. Conclusion Uridine diphosphate galactose can protect the chilled rabbit platelets,prolong the circulation of refrigerated rabbit platelets,and do not impair the chilled rabbit platelet function.