Beutler E, Kuhl W, West C
Blood. 1982 Jun;59(6):1141-7.
Although it is recognized that red cells lose membrane during storage, estimation of the osmotic fragility of erythrocytes has not previously proven to be a useful measurement of the storage lesion. Erythrocytes from blood stored in CPD-A2 were found to have a markedly increased osmotic fragility. A major portion of this increase was found to be due to accumulation of lactate, which is only slowly transported from within erythrocytes and which therefore exerts a strong osmotic effect in the usual osmotic fragility test. After an hour's incubation in a large volume of iso-osmotic buffer, the osmotic fragility curve of stored erythrocytes was much more nearly normal. Such cells were found to have a volume 5%--8% greater than that of normal cells, indicating that even after removal of lactate more osmotically active material was present in the stored erythrocytes than in fresh cells. Most of this differences can be accounted for by substitution of chloride ion for 2,3-DPG, since chloride exerts approximately 3.7 times the osmotic effect of 2,3-DPG per unit charge. In addition to the shift in osmotic fragility produced by the increased intracellular osmotically active material, a "fragile tail" of red cells was also present. Stored erythrocytes were labeled with 51Cr and reinfused into the volunteer donors. The osmotic fragility of the reinfused cells was estimated using a technique of sequential osmotic hemolysis that permitted accurate estimation of osmotic fragility of transfused cells using very small amounts of 51Cr. The osmotic fragility of the reinfused cells became less than those of fresh cells after 24 hr and was exactly the same as those of fresh cells after 4 days. The fragile tail disappeared at a rate that approximated the rate of loss of nonviable erythrocytes from the circulation as measured by 51Cr. These findings are consistent with the preferential destruction of a subpopulation of red cells with a diminished surface area.
尽管人们认识到红细胞在储存过程中会丢失膜,但红细胞渗透脆性的评估此前尚未被证明是一种用于衡量储存损伤的有效方法。发现储存在CPD - A2中的血液中的红细胞渗透脆性显著增加。发现这种增加的主要部分是由于乳酸的积累,乳酸从红细胞内转运缓慢,因此在通常的渗透脆性试验中会产生强烈的渗透效应。在大量等渗缓冲液中孵育一小时后,储存红细胞的渗透脆性曲线更接近正常。发现这些细胞的体积比正常细胞大5% - 8%,这表明即使去除乳酸后,储存红细胞中仍比新鲜细胞含有更多具有渗透活性的物质。这种差异的大部分可以用氯离子取代2,3 - 二磷酸甘油酸(2,3 - DPG)来解释,因为每单位电荷氯离子产生的渗透效应约为2,3 - DPG的3.7倍。除了细胞内渗透活性物质增加导致的渗透脆性变化外,还存在红细胞的“脆弱尾”。用51Cr标记储存的红细胞并重新注入志愿者供体体内。使用连续渗透溶血技术估计重新注入细胞的渗透脆性,该技术允许使用极少量的51Cr准确估计输血细胞的渗透脆性。重新注入细胞的渗透脆性在24小时后变得低于新鲜细胞,4天后与新鲜细胞完全相同。脆弱尾消失的速率近似于通过51Cr测量的循环中无活力红细胞丢失的速率。这些发现与表面积减小的红细胞亚群优先被破坏一致。