Red blood cells ageing markers: a multi-parametric analysis
Red blood cells ageing markers: a multi-parametric analysis
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DOI：
10.2450/2017.0318-16
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发表时间：
2017-01-01
影响因子：
3.7
通讯作者：
Prudent, Michel
中科院分区：
文献类型：
--
作者：
Bardyn, Manon;Rappaz, Benjamin;Prudent, Michel
背景在柠檬酸盐-磷酸盐-葡萄糖中收集的红细胞可在4 ℃下在盐水-腺嘌呤-葡萄糖-甘露醇添加剂溶液中储存长达42天。在这种受控但仍然是人工的离体老化期间，红细胞积累损伤，这些损伤在输血时可以是可逆的或不可逆的。本研究的目的是跟踪几个参数反映细胞代谢，抗氧化防御，形态和膜动力学在存储过程中。在71天内，每周跟踪5次红细胞浓缩物。细胞外葡萄糖和乳酸浓度，总抗氧化能力，以及减少和氧化的细胞内谷胱甘肽水平进行了定量。还评价了微泡形成、溶血百分比和血液学参数。最后，采用免标记数字全息显微镜观察细胞形态学变化和细胞膜波动。抗氧化能力以及细胞内谷胱甘肽浓度首先增加，分别在一周和两周后达到最大值。第5周出现不可逆的形态学病变，椎间盘细胞开始转化为短暂的棘细胞，最后是球形细胞。与此同时，微泡和溶血开始呈指数级上升。六周后（有效期），细胞内谷胱甘肽减少了25%，反映了氧化应激增加。在从盘状细胞向球形细胞的形态转变过程中，细胞膜的波动幅度降低。在储存期间，各种类型的病变在不同的化学和细胞水平上积累，这可能影响它们在输血后的体内恢复。储存四周后观察到显著效果，这证实了最近的临床数据。延长的随访期允许捕获深部储存病变。有趣的是，如前所述，变化的严重程度在捐助者之间存在差异。
Background. Red blood cells collected in citrate-phosphate-dextrose can be stored for up to 42 days at 4 degrees C in saline-adenine-glucose-mannitol additive solution. During this controlled, but nevertheless artificial, ex vivo ageing, red blood cells accumulate lesions that can be reversible or irreversible upon transfusion. The aim of the present study is to follow several parameters reflecting cell metabolism, antioxidant defences, morphology and membrane dynamics during storage.Materials and methods. Five erythrocyte concentrates were followed weekly during 71 days. Extracellular glucose and lactate concentrations, total antioxidant power, as well as reduced and oxidised intracellular glutathione levels were quantified. Microvesiculation, percentage of haemolysis and haematologic parameters were also evaluated. Finally, morphological changes and membrane fluctuations were recorded using label-free digital holographic microscopy.Results. The antioxidant power as well as the intracellular glutathione concentration first increased, reaching maximal values after one and two weeks, respectively. Irreversible morphological lesions appeared during week 5, where discocytes began to transform into transient echinocytes and finally spherocytes. At the same time, the microvesiculation and haemolysis started to rise exponentially. After six weeks (expiration date), intracellular glutathione was reduced by 25%, reflecting increasing oxidative stress. The membrane fluctuations showed decreased amplitudes during shape transition from discocytes to spherocytes.Discussion. Various types of lesions accumulated at different chemical and cellular levels during storage, which could impact their in vivo recovery after transfusion. A marked effect was observed after four weeks of storage, which corroborates recent clinical data. The prolonged follow-up period allowed the capture of deep storage lesions. Interestingly, and as previously described, the severity of the changes differed among donors.