海藻糖水替代机制对冻干红细胞保护作用的探讨

The protective impact of trehalose on lyophilized red blood cells

目的探讨海藻糖对冻干红细胞复水回收率和残余水含量的影响及其对红细胞膜的保护机制。方法将红细胞分别与100、200、400、600、800、1 000 mmol/L海藻糖负载溶液孵育,共6组,并设置对照组,用海藻糖检测试剂盒特异性检测红细胞中海藻糖加载量;用冷冻干燥机对红细胞做冷冻干燥,Karl-Fisher滴定法结合卡式炉法测定冻干红细胞中残余水含量;细胞计数仪检测复水后红细胞数目。结果随着负载海藻糖溶液浓度的上升,冻干红细胞复水回收率先平稳后上升,当负载溶液浓度达到800 mmol/L时回收率最高;残余水含量先保持不变,然后下降,当负载溶液浓度达到800和1 000 mmol/L时,残余水含量（%）3.47±0.21和2.43±0.18。此外,冻干红细胞组与冻干血影细胞组残余水相同。结论初步证实了海藻糖通过水替代作用保护红细胞膜。

Objective To investigate the protective mechanism of trehalose on the cell membrane of lyophilized red blood cells（ RBCs） by exploring its influence on the recovery rate and water residuals. Methods RBCs were incubated with various concentration of trehalose solution（ 100、200、400、600、800、1 000 mmol/L） and control group were prepared according to conventional methods. Their trehalose content were determined by Trehalose kits. Cells loaded with trehalose were lyophilized with a freeze-dryer. Their residual water content were determined by Karl-Fisher titration and the recovery rates were analyzed with automatic blood cell counter. Results As the concentration of trehalose increased,recovery rates of RBCs kept steady first and then increased; The highest recovery rate is obtained at the concentration of 800 m M. In terms of residual water content,it presents a generally decreasing trend as the the concentration of trehalose increased. The figures for residual water dropped sharply to 3. 47 ± 0. 21% and 2. 43 ± 0. 18% when the concentration reaches to 800 mmol/L and 1 000 mmol/L,respectively. Conclusion Our experiments primarily confirm that the protective mechanism of trehalose on lyophilized red blood cells was water replacement.