Effect of a glycerol-containing hypotonic medium on erythrocyte phospholipid asymmetry and aminophospholipid transport during storage.
Effect of a glycerol-containing hypotonic medium on erythrocyte phospholipid asymmetry and aminophospholipid transport during storage.
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含甘油的低渗介质对储存期间红细胞磷脂不对称性和氨基磷脂转运的影响。
DOI：
10.1016/s0005-2736(97)00157-0
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发表时间：
1997
期刊：
影响因子：
--
通讯作者：
Daleke,DL
中科院分区：
文献类型：
--
作者：
Dumaswala,UJ;Wilson,MJ;José,T;Daleke,DL
我们实验室之前的研究表明，在血库储存条件下，含甘油的低渗实验添加剂溶液（EAS 25）比传统储存介质Adsola能更好地维持红细胞（RBC） ATP和脂质含量。本研究的目的是通过测量跨膜磷脂不对称性和储存红细胞的膜完整性来确定EAS 25的保护作用机制。分离的红细胞储存在EAS 25或Adsol®中。在0、42和84天后分析红细胞，并在84天后分析红细胞脱落的囊泡。磷脂不对称通过磷脂酶a2消化（红细胞）和凝血酶原复合物（红细胞，囊泡）的激活来测定。与EAS 25相比，Adsol®储存后红细胞膜的磷脂酰乙醇胺外化量显著增加（P < 0.01）（分别为44.3%±11.7比25.3%±5.7）。红细胞凝血酶原转化活性显著低于脱落囊泡（P < 0.001），表明磷脂酰丝氨酸存在于囊泡的外层，而不存在于红细胞膜。在两种培养基中，红细胞内定向的氨基磷脂转运率下降了约50%，谷胱甘肽水平下降了约50%。EAS 25贮藏红细胞中红细胞胆固醇和磷脂含量显著高于Adsole （P < 0.01）。结果表明，尽管氨基酸磷脂转运速率和GSH浓度降低，但在EAS 25中比在Adsol®中更好地保持了红细胞磷脂的不对称性。数据表明，低渗EAS中的甘油有助于在储存过程中保持红细胞脂质组织和膜完整性。
Previous studies from our laboratory have shown that under blood bank storage conditions red blood cell (RBC) ATP and lipid content were better maintained in a glycerol-containing hypotonic experimental additive solution (EAS 25) than in the conventional storage medium Adsola. The objective of this study was to determine the mechanism of the protective effect of EAS 25, by measuring transmembrane phospholipid asymmetry and the membrane integrity of stored RBCs. Split units of packed RBCs were stored in either EAS 25 or Adsol®. RBCs were analyzed after 0, 42, and 84 days and vesicles shed from stored RBCs were analyzed after 84 days of storage. Phospholipid asymmetry was measured by phospholipase A2digestion (RBCs) and activation of the prothrombinase complex (RBCs, vesicles). RBC membrane exhibited a significantly greater (P < 0.01) amount of phosphatidylethanolamine externalized after storage in Adsol® than in EAS 25 (44.3% ± 11.7 vs. 25.3% ± 5.7, respectively). Prothrombin converting activities in RBCs were significantly lower than in shed vesicles (P < 0.001) suggesting the presence of phosphatidylserine in the outer monolayer of vesicle, but not in RBC membranes. The rates of inwardly-directed aminophospholipid transport in RBCs decreased by ∼ 50% and glutathione levels decreased by ≈ 50% in both media. RBC cholesterol and phospholipid content of stored RBCs remained significantly greater (P > 0.01) in EAS 25 than in Adsole. The results indicate that despite comparable reduction in the rate of aminophospholipid transport and reduced GSH concentrations, RBC phospholipid asymmetry was better maintained during storage in EAS 25 than in Adsol®. The data suggest that glycerol in the hypotonic EAS helps preserve RBC lipid organization and membrane integrity during storage.