Dumaswala U J, Dumaswala R U, Levin D S, Greenwalt T J
Hoxworth Blood Center, University of Cincinnati Medical Center, OH 45267-0055, USA.
Blood. 1996 Feb 15;87(4):1612-6.
In earlier studies we have shown that a final concentration of 0.69% glycerol in blood mixed with an experimental additive solution, EAS 25, improves the in vitro quality and in vivo survival of red blood cells (RBCs). The objective of this study was to determine if the better preservation of RBCs in EAS 25 is correlated with the improved maintenance of membrane lipids and proteins and decreased vesiculation. Split units of RBCs were stored in Adsol or EAS 25 (mmol/L: adenine 2/2, dextrose 122/110, mannitol 42/55, glycerol 0/150, NaCl 154/50). After 12 weeks storage, RBC and microvesicle membranes were analyzed for cholesterol, phospholipid, diphenyl hexatriene fluorescence anisotropy, and acetylcholinesterase (AchE) activity. Bands 3 and 4.1 were identified in the microvesicle membranes by immunoblotting. The RBC membrane cholesterol, phospholipids, and AchE remained higher in EAS 25 than in Adsol (P < .001). Vesicle membrane lipids and AchE in EAS 25 were significantly less than in Adsol (P < .001). The fluidity of stored cells in both the solutions was greater than the prestorage samples. Immunoblotting analyses showed that bands 3 and 4.1 were greatly reduced in the microvesicle membranes shed by the RBCs stored in EAS 25 compared with those formed in Adsol.
在早期研究中，我们已经表明，血液中与实验添加剂溶液EAS 25混合的甘油终浓度为0.69%时，可改善红细胞（RBC）的体外质量和体内存活率。本研究的目的是确定在EAS 25中红细胞保存效果更好是否与膜脂质和蛋白质的维持改善以及囊泡形成减少相关。将红细胞的分离单位储存在Adsol或EAS 25中（毫摩尔/升：腺嘌呤2/2、葡萄糖122/110、甘露醇42/55、甘油0/150、氯化钠154/50）。储存12周后，分析红细胞和微囊泡膜中的胆固醇、磷脂、二苯基己三烯荧光各向异性和乙酰胆碱酯酶（AchE）活性。通过免疫印迹法在微囊泡膜中鉴定出带3和带4.1。EAS 25中红细胞膜胆固醇、磷脂和AchE仍高于Adsol（P <.001）。EAS 25中囊泡膜脂质和AchE明显少于Adsol（P <.001）。两种溶液中储存细胞的流动性均大于储存前样品。免疫印迹分析表明，与在Adsol中形成的微囊泡膜相比，在EAS 25中储存的红细胞脱落的微囊泡膜中带3和带4.1大大减少。
