冻干血小板聚集和活化的实验研究

Experimental study on the aggregation and activation of the freeze-dried platelets

目的了解血小板在冻干前、后功能活性的变化。方法应用流式细胞仪检测冻干复水(再水化)血小板及未冷冻干燥血小板胞膜CD61、CD62p、PAC-1的分子表达,评价冻干复水后血小板活化状态;应用血小板聚集仪检测通过诱导剂瑞斯托霉素、二磷酸腺苷诱导的血小板最大聚集率,分析冻干血小板复水后血小板聚集率变化。结果血小板冻干保护剂渗透压值为(2 055.6±123.8)m OSmol/kg;冻干前及复水后血小板分布宽度(PDW)(%)分别为15.50±3.05、18.29±0.55,MPV(f L)分别为9.01±1.84、8.63±0.58;冻干血小板复水回收率(%)为81.57±12.57;冷冻前及复水后血小板最大聚集率(%)分别为34.30±33.14、22.10±23.25;冷冻前及冻干复水后血小板胞膜CD61分子表达(%)分别为43.17±13.55、48.64±13.24,CD62p分子表达(%)分别为17.34±6.47、9.79±4.48,PAC-1分子表达(%)分别为4.92±6.32、4.22±4.64。结论冷冻前及复水后血小板最大聚集率的变化甚微,冻干复水后血小板仍具仍具有一定的存活能力;冷冻前及冻干复水后血小板胞膜CD61、CD62p和PAC-1分子表达略有变化(P>0.05)。

Objective To understand the changes in the function and activity of the.platelets before and after freeze-dr- ying. Methods The expressions of the CD61,62p and PAC-1 of the platelets before freeze-drying and rehydration were analyzed by flow cytometry （FCM）. The state of the platelets after freeze-drying was evaluaied. Through Ristocetin and ADP inducing agent, the maximum aggregation rates of the no-induced platelets ＇and induced platelets were detected by platelet aggregation instrument. The changes in aggregation rate of the platelets after freeze-drying were analyzed. Results The osmotic pressure value of the freeze-drying protective agent of the platelets was （2 055.6 ± 123. 8） mOSMOL/KG. The PDW （%） results of the platelets before lyophilization and rehydration were 15. 50 ± 3.05 and 18.29 ± 0. 55. The MPV （fL） values of the platelets before lyophilization and rehydration were 9. 01±1.84 and 8.63 ±0. 58. The recovery rate （%） of the freezedried platelets was 81.57 ± 12. 57. The maximum aggregation rates （ % ） of the platelets before lyophilization and rehydration were 34. 30 ±33. 14 and 22. 10 ±23.25. The results （%） of CD61 expression of the platelets before lyophilization and rehydration were 43.17 ± 13.55 and 48.64 ±13.24. The results （ % ） of CD62p expression of the platelets before lyophilization and rehydration were 17.34 ±6. 47 and 9.79 ± 48. The results （ % ） of PAC-1 expression of the platelets before lyophilization and rehydration were 4.92 ± 6. 32 and 4.22 ±4. 64. Conclusion The maximum aggregation rate of the platelets before lyophilization has shown no Significant difference when compared with that of the rehydrated platelets： The lyophilized platelets still have a certain ability of survival. The expression of CD61, CD62p and PAC-1 slightly changes in the platelets before lyophilization and rehydration.