Red blood cell washing, nitrite therapy, and antiheme therapies prevent stored red blood cell toxicity after trauma-hemorrhage.
Red blood cell washing, nitrite therapy, and antiheme therapies prevent stored red blood cell toxicity after trauma-hemorrhage.
复制标题
红细胞清洗、亚硝酸盐疗法和抗血红素疗法可预防创伤出血后储存的红细胞中毒。
DOI：
10.1016/j.freeradbiomed.2015.04.025
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发表时间：
2015-08
影响因子：
7.4
通讯作者：
Patel RP
中科院分区：
文献类型：
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作者：
Stapley R;Rodriguez C;Oh JY;Honavar J;Brandon A;Wagener BM;Marques MB;Weinberg JA;Kerby JD;Pittet JF;Patel RP
储存的红细胞（RBC）的输注与创伤患者的发病率和死亡率增加有关。储存的红细胞的促氧化、促炎和一氧化氮 (NO) 清除特性被认为是这种关联的基础。在这项研究中，我们确定了红细胞清洗、亚硝酸盐和抗血红素治疗对创伤引起的出血情况下储存的红细胞依赖性毒性的影响。使用小鼠（C57bl/6）创伤出血和复苏模型，其中保存 1 或 3 单位红细胞 0-10 天。测试的变量包括是否清洗红细胞以去除清除 NO 的较低 MWt 成分、使用亚硝酸盐进行 NO 补充治疗或通过血红素清除减轻游离血红素毒性或防止 TLR4 激活。通过评估急性肺损伤指数（气道水肿和炎症）和生存来确定储存的红细胞毒性。输注 5 天红细胞会增加以 BAL 蛋白和中性粒细胞积累为指标的急性肺损伤。输血前清洗 5 天红细胞并不能减少这种损伤，而亚硝酸盐治疗则可以。输注 10 天红细胞会引起更严重的损伤，导致约 90% 的致死率，而输注 5 天红细胞则<15%。清洗和亚硝酸盐疗法均能显着防止 10 天红细胞引起的致死，这表明当损伤刺激更严重时，清洗可能具有保护作用。最后，开发了光谱反卷积测定法来同时测量储存的红细胞上清液中的游离血红素和血红蛋白，这表明储存的人和小鼠红细胞中的游离血红素和血红蛋白均显着增加。输注游离血红素部分再现了储存红细胞介导的毒性。此外，使用 TAK-242 抑制血红素刺激的 TLR4 信号传导或血红素依赖性游离血红素隔离可显着防止 5 天和 10 天小鼠红细胞依赖性毒性。这些数据表明红细胞清洗、亚硝酸盐治疗和/或抗血红素和 TLR4 策略可以预防储存的红细胞毒性。
Transfusion of stored red blood cells (RBCs) is associated with increased morbidity and mortality in trauma patients. Pro-oxidant, pro-inflammatory and nitric oxide (NO) scavenging properties of stored RBC are thought to underlie this association. In this study we determined the effects of RBC washing, nitrite and anti-heme therapy on stored RBC-dependent toxicity in the setting of trauma-induced hemorrhage. A murine (C57bl/6) model of trauma-hemorrhage and resuscitation with 1 or 3 units of RBC stored for 0–10d was used. Tested variables included whether washing RBC to remove lower MWt components that scavenge NO, NO-repletion therapy using nitrite or mitigation of free heme-toxicity by heme scavenging or preventing TLR4 activation. Stored RBC toxicity was determined by assessment of acute lung injury indices (airway edema and inflammation) and survival. Transfusion with 5d RBC increased acute lung injury indexed by BAL protein and neutrophil accumulation. Washing 5d RBC prior to transfusion did not decrease this injury, whereas nitrite therapy did. Transfusion with 10d RBC elicited a more severe injury resulting in ~90% lethality, compared to <15% with 5d RBC. Both washing and nitrite therapy significantly protected against 10d RBC-induced lethality, suggesting that washing may be protective when the injury stimulus is more severe. Finally, a spectral deconvolution assay was developed to simultaneously measure free heme and hemoglobin in stored RBC supernatants, which demonstrated significant increases of both in stored human and mouse RBC. Transfusion with free heme partially recapitulated the toxicity mediated by stored RBC. Furthermore, inhibition of TLR4 signaling, which is stimulated by heme, using TAK-242, or hemopexin-dependent sequestration of free heme significantly protected against both 5d and 10d mouse RBC-dependent toxicity. These data suggest that RBC washing, nitrite therapy and / or anti-heme and TLR4 strategies may prevent stored RBC toxicities.