红细胞保存期对冻干前海藻糖负载量的影响研究

Study of Shelf Life of Red Blood Cells Effecting Intracellular Trehalose

探讨冻干前红细胞保存期对海藻糖负载量的影响,确定红细胞负载海藻糖的最佳条件。在37℃条件下,4℃保存0、24、48和72 h的红细胞在海藻糖浓度800 mmol/L的负载缓冲液中孵育7 h后,检测胞内海藻糖浓度和胞外外游离血红蛋白浓度。经相同条件孵育后,4组红细胞对海藻糖的摄取量基本相同,分别为(49.747±2.253)、(50.316±0.612)、(49.768±2.179)和(49.013±1.991)mmol/L,各组之间无统计学差异。4℃保存24、48和72 h的红细胞相对于新鲜红细胞,3组负载红细胞胞外游离血红蛋白明显增加,分别高达(8.473±2.138)g/L(P<0.05)、(12.697±1.787)g/L(P<0.01)和(14.036±2.796)g/L(P<0.01)。说明冻干前红细胞4℃保存期不会影响海藻糖负载量,红细胞负载海藻糖的主要影响因素仍为负载温度、负载时间和负载缓冲液中海藻糖浓度,若固定孵育条件,红细胞对海藻糖的胞内负载量基本保持不变。但随保存期的延长,红细胞在高渗环境中孵育更易损伤,溶血率明显增加。所以,为达到更好的海藻糖负载效果,应尽量采用新鲜红细胞进行处理。

To study whether the shelf life of red blood cells before freeze-drying can effect trehalose loading, to determine the best conditions for red blood cell loading trehalose. At 37℃ condition, red blood cells preserved in 4 ℃ for 0,24,48 and 72 h were incubated in the loading solution which trehalose concentration is 800 mmol / L for 7 h, then detected the intracellular trehalose and extracellular concentrations of free hemoglobin. After incubated in the same conditions, the four groups of trehalose intake of red blood cell is basically the same, respectively, （49.747± 2.253）,（50.316 ± 0.612）,（49.768 ± 2.179）and（49.013± 1.991）mmol/ L, no significant difference among the groups. Red blood cells preserved in 4 ℃ for 0,24,48 and 72 h compared to fresh red blood ceils, extracellular free hemoglobin in these three groups significantly increased, were as high as（8. 473± 2. 138 ） g/L（P〈0.05）,（12.697 ± 1.787）g/L（P〈0.01 ）, and（14.036 ± 2. 796）g/L（P〈0.01）. It is concluded that the main factor effecting intracellular trehalose in red blood cell was loading temperature, loading time and the trehalose concentration in load buffer, if fixig incubation conditions, intracellular trehalose in red blood ceils remained basically unchanged, there was nothing to do with the shelf life of red blood cells before freeze-drying. However, with the extension of shelf life, red blood cells incubated in the hypertonic environment, more easily damaged, with a marked increase in the rate of hemolysis. Therefore, in order to achieve a better effect of trehalose loading, we should use fresh red blood cells for processing.