二甲亚砜与第二信使效应剂对深低温冻存血小板膜CD42b、CD62p表达的影响

EFFECTS OF DIMETHYL SULFOXIDE AND SECOND-MESSENGER EFFECTOR ON CRYOPRESERVED PLATELET MEMBRANE EXPRESSIONS CD42b AND CD62p

①目的观察体积分数0.02二甲亚砜(DMSO)联合第二信使效应剂(ThromboSol,TS)在-196 ℃时对浓缩血小板膜CD42b、CD62p表达的影响,并了解体积分数0.02 DMSO+TS对浓缩血小板的长期(6个月)保存效果.②方法分别应用体积分数0.02 DMSO、体积分数0.06 DMSO、体积分数0.02 DMSO+TS作为冻存液-196 ℃冷冻保存浓缩血小板,并在冻存后1、3及6个月时分别取出复温,流式细胞仪检查血小板膜CD42b、CD62p表达水平.③结果冻存后1、3及6个月,体积分数0.02 DMSO+TS组与体积分数0.06 DMSO组血小板CD42b的表达水平差异无显著性(F=3.13～5.01,q=1.12～1.76,P＞0.05),两者均高于体积分数0.02 DMSO组(q=3.17～4.06,P＜0.05);但与新鲜组相比,3组冷冻保存血小板CD42b表达水平均有所下降(q=3.09～4.15,P＜0.05).体积分数0.02 DMSO+TS组CD62p的表达水平低于体积分数0.06 DMSO组和体积分数0.02 DMSO组,差异有显著性(F=5.75～9.77,q=3.20～4.34,P＜0.05),后两组差异无统计学意义(q=1.55～2.02,P＞0.05);与新鲜组相比,3组均明显升高,差异有显著性(q=4.85～7.35,P＜0.01).④结论第二信使效应剂的应用降低了DMSO的使用浓度,在抑制血小板的体外激活方面,体积分数0.02 DMSO+TS的效果优于体积分数0.06 DMSO,且长期(6个月)保存效果稳定.

Objective To observe the effect of cryopreservation with 0.02 dimethyl sulfoxide(DMSO) plus ThromboSol, a mixture of second-messenger effectors, on platelet membrane CD42b and CD62p, and to investigate the long-term (6 months) preservation effect of the platelet storage solution. Methods Platelet concentrates were treated with 0.02 DMSO, 0.06 DMSO and 0.02 DMSO plus ThromboSol, and then placed in liquid nitrogen. The samples were thawed and expression levels of CD42b and CD62p on platelet membrane were analyzed after one month, three months and six months, respectively. Results The platelet expression level of CD42b in the 0.02 DMSO-plus-ThromboSol group was not significantly different from those in the 0.06 DMSO group (F=3.13-5.01,q=1.12-1.76, P>0.05), and the levels of both groups were higher than those in the 0.02 DMSO group (q=3.17-4.06, P<0.05) one, three and six months later. The CD42b levels of the three groups were lower than those of the fresh platelets (q=3.09-4.15, P<0.05). The platelets treated with 0.02 DMSO plus ThromboSol showed statistically lower CD62p expression than those frozen with 0.06 DMSO and 0.02 DMSO(F=5.75-9.77,q=3.20-4.34,P<0.05). There were no obvious differences in CD62p expression between platelets treated with 0.06 DMSO and 0.02 DMSO (q=1.55-2.02, P>0.05). But the three groups of frozen platelets showed significant increase of surface expression of CD62p in comparison to the fresh platelets (F=3.95, q=4.85-7.35, P<0.01). Conclusion The second-messenger effectors could reduce the DMSO concentration. The 0.02 DMSO plus ThromboSol is superior to 0.06 DMSO in inhibiting the activation of platelets, and is stable within six months.