摘要
目的探讨不同复水条件对冻干血小板功能的影响。方法经过添加激活抑制剂(PGE1、左旋精氨酸、植酸钠和百维利肽)、DMSO和海藻糖等低温保护剂、冷冻干燥后获得冻干血小板,在一系列不同复水程序下对冻干血小板复水,应用流式仪检测并分析其CD62p和PAC-1的表达,评价血小板活化状态。通过正交设计,研究不同复水程序包括复水溶液、复水温度、复水方式3个影响因素对冻干血小板复水后活性的影响。结果 3个对冻干血小板复水后活化的影响因素排序:复水温度>复水方式>复水液(P<0.05);37℃条件下冻干血小板复水后血小板膜蛋白CD62p和PAC-1活化率最低,分别为(5.96±2.51)%和(4.55±1.97)%;预水化后再行四步添加复水可降低冻干血小板膜蛋白CD62p和PAC-1的表达率至(6.68±2.62)%和(5.45±2.06)%;添加聚乙烯吡咯烷酮(PVP)和血小板抑制剂的2号复水液对冻干血小板复水后膜蛋白CD62p和PAC-1的表达率影响最低,分别为(6.72±3.35)%和(5.28±2.30)%。总体模型评价进一步显示,第5实验组,即在37℃条件下加入PVP和血小板抑制剂的复水液配方,先预水化后再行四步法添加复水,其复水血小板活化率最低,CD62p和PAC-1的表达率分别为(3.06±1.36)%和(2.70±0.84)%。结论 37℃下,饱和蒸汽环境中预水化后,采用加入PVP及血小板抑制剂的复水液四步添加复水,其复水血小板活化率最低。
Objective To explore the effect of different rehydration conditions on function of the lyophilized plate- lets. Methods After the lyophilization, platelets were given different rehydration conditions including different formula of rehydration solution, the rehydration temperature and the addictive method of rehydration solution using the orthogonal de- sign. The expression of CD62p and PAC-1 were measured as the indexes of platelet activation. Results The impact of 3 factors from large to small order on viability of freeze-dried platelets was rehydration temperature, addictive method of rehydration solution and formular of rehydration solution, which all showed significantly different （ P 〈 0. 05 ）. If freeze-dried platelets were rehydrated under 37℃ , the expression of membrane protein CD62p and PAC-1 was the lowest, which was （5.96±2. 51 ） % and （4. 55 ± 1.97） % , respectively. If the freeze-dried platelets were rehydrated by the vapor first and then rehydration solution by 4 steps, the expression of platelet membrane protein CD62p and PAC-1 was also reduced, which was （6. 68 ± 2. 62 ） % and （5.45 ± 2. 06 ） %, respectively. If freeze-dried platelets were rehydrated with the second rehydration solution with polyvinylpyrrolidone （PVP） and platelet inhibitors,the expression of membrane protein CD62p and PAC- 1 was the lowest, which was （ 6.72 ± 3.35 ） % and （ 5.28± 2.30） %, respectively. Tile overall evaluation model also showed that the 5th experimental groups of freeze-dried platelets was optimal whose procedure was the rehydration solution with PVP, and platelet inhibitors and rehydrated by the vapor first, and then rehydration solution by 4 steps under 37 ℃. After rehydration, the activation of platelets was the lowest, with the expression of CD62p and PAC-1 of （3. 06 ± 1.36）% and （2. 70 ± 0. 84）%, respectively. Conclusion The experimental results showed that with preliminary hydration in 37℃sat- urated steam environment, using rehydration solution combin
出处
《中国输血杂志》
CAS
CSCD
北大核心
2013年第4期338-342,共5页
Chinese Journal of Blood Transfusion
基金
国家自然科学基金(30271443)
关键词
血小板
冻干
复水
血小板活化
血小板抑制剂
platelet
lyophilization
rehydration
platelet activation
platelet inhibitor
