Haematology and Haemotherapy Hospital Foundation of Amazonas, Manaus, Amazonas.
Faculty of Pharmaceutical Sciences, Federal University of Amazonas, Manaus, Amazonas.
Blood Transfus. 2021 Jul;19(4):309-316. doi: 10.2450/2020.0062-20. Epub 2020 Nov 3.
Red blood cells (RBC) are subject to oxidative stress by reactive oxygen species (ROS) during storage. Molecular characterisation of oxidative stress in stored RBC, which may also occur in other blood components during long periods of storage, is rare.
Our study included 45 healthy RBC donors recruited in Brazil. Blood was collected into standard Grifols Triple Bags containing CPD SAG-M. Haematological values, biochemical data, and oxidative stress markers were assessed weekly during storage until 42 days after collection. GSTM1 and GSTT1 were determined by multiplex-polymerase chain reaction (PCR), while GSTP1 rs1695 and rs1871042, CAT rs1001179, and SOD2 rs4880 were evaluated by real-time PCR.
A direct proportional relationship was found between storage time and levels of ROS and thiobarbituric acid reactive substances (TBARS, indicators of lipid peroxidation) (p<0.001). These parameters were indirectly proportional to ABTS values (p<0.001). The plasma concentration of TBARS was associated with GSTP1 303, GSTP1 -16, and SOD2 47 genotypes. Single-nucleotide polymorphisms at the CAT C-262T gene were not associated with TBARS, nor were oxidative markers of ROS.
Prolonged storage may result in the onset of erythrocyte deterioration. Our results clearly indicate that erythrocytes are capable of attenuating ROS for 2 weeks of storage. We observed an association between elevated TBARS levels and the presence of GSTP1 and SOD2 variants in stored RBC. Although notable for heterozygous variants, this association was even stronger for the homozygous variants GSTP1 rs1695 (303), GSTP1 rs1871042 (-16), and SOD2 rs4880 (47). These findings accentuate the importance of genetic factors in storage lesions and will expand our understanding and consideration of endogenous and exogenous causes in improving clinical treatment with blood transfusions.
在储存过程中，红细胞（RBC）会受到活性氧（ROS）引起的氧化应激。在储存过程中，其他血液成分也可能发生长期储存的氧化应激，对其进行分子特征分析的情况很少见。
我们的研究包括 45 名在巴西招募的健康 RBC 供体。血液采集到含有 CPD SAG-M 的标准 Grifols Triple 袋中。在储存期间，每周评估血液学值、生化数据和氧化应激标志物，直到采集后 42 天。使用多重聚合酶链反应（PCR）测定 GSTM1 和 GSTT1，而使用实时 PCR 评估 GSTP1 rs1695 和 rs1871042、CAT rs1001179 和 SOD2 rs4880。
发现储存时间与 ROS 和硫代巴比妥酸反应物质（TBARS，脂质过氧化的指标）水平之间存在直接比例关系（p<0.001）。这些参数与 ABTS 值呈间接比例关系（p<0.001）。血浆 TBARS 浓度与 GSTP1 303、GSTP1 -16 和 SOD2 47 基因型相关。CAT C-262T 基因的单核苷酸多态性与 TBARS 或 ROS 的氧化标志物无关。
长时间储存可能导致红细胞恶化。我们的结果清楚地表明，红细胞能够在储存的前 2 周内减轻 ROS。我们观察到储存 RBC 中升高的 TBARS 水平与 GSTP1 和 SOD2 变体的存在之间存在关联。尽管杂合变体明显，但 GSTP1 rs1695（303）、GSTP1 rs1871042（-16）和 SOD2 rs4880（47）的纯合变体的相关性更强。这些发现强调了遗传因素在储存损伤中的重要性，并将扩展我们对改善输血临床治疗的内源性和外源性原因的理解和考虑。
