红细胞衰老标志物：多参数分析
Red blood cells ageing markers: a multi-parametric analysis.
作者信息
Bardyn Manon, Rappaz Benjamin, Jaferzadeh Keyvan, Crettaz David, Tissot Jean-Daniel, Moon Inkyu, Turcatti Gerardo, Lion Niels, Prudent Michel
机构信息
Blood Products Research Laboratory, Interregional Blood Transfusion SRC, Epalinges, Switzerland.
Swiss Federal Institute of Technology, Biomolecular Screening Facility, School of Life Sciences, Lausanne, Switzerland.
出版信息
Blood Transfus. 2017 May;15(3):239-248. doi: 10.2450/2017.0318-16.
BACKGROUND
Red blood cells collected in citrate-phosphate-dextrose can be stored for up to 42 days at 4 °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.
背景
采集于枸橼酸盐 - 磷酸盐 - 葡萄糖溶液中的红细胞，在添加了生理盐水 - 腺嘌呤 - 葡萄糖 - 甘露醇的溶液中于4℃可储存长达42天。在这种可控但仍是人为的体外老化过程中，红细胞会积累一些病变，这些病变在输血后可能是可逆的或不可逆的。本研究的目的是在储存过程中跟踪反映细胞代谢、抗氧化防御、形态和膜动力学的几个参数。
材料与方法
在71天内每周对五份红细胞浓缩液进行跟踪。对细胞外葡萄糖和乳酸浓度、总抗氧化能力以及细胞内还原型和氧化型谷胱甘肽水平进行定量分析。还评估了微囊泡形成、溶血百分比和血液学参数。最后，使用无标记数字全息显微镜记录形态变化和膜波动情况。
结果
抗氧化能力以及细胞内谷胱甘肽浓度首先升高，分别在第1周和第2周后达到最大值。在第5周出现不可逆的形态病变，此时双凹圆盘状红细胞开始转变为过渡性棘状红细胞，最终变为球形红细胞。与此同时，微囊泡形成和溶血开始呈指数上升。在六周（有效期）后，细胞内谷胱甘肽减少了25%，反映出氧化应激增加。在从双凹圆盘状红细胞向球形红细胞的形态转变过程中，膜波动的幅度减小。
讨论
在储存过程中，不同化学和细胞水平上积累了各种类型的病变，这可能会影响它们输血后的体内恢复情况。在储存四周后观察到显著影响，这与最近的临床数据相符。延长的随访期使得能够捕捉到深度储存病变。有趣的是，如先前所述，不同供体之间变化的严重程度有所不同。
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