Division of Traumatology, Department of Surgery, Critical Care and Acute Care Surgery, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA.
J Surg Res. 2013 Sep;184(1):422-9. doi: 10.1016/j.jss.2013.05.097. Epub 2013 Jun 19.
Hemorrhagic shock is a leading cause of death following severe trauma, and platelet transfusions are frequently necessary to achieve hemostasis. Platelets, however, require special storage conditions, and storage time has been associated with loss of platelet quality. We hypothesized that standard storage conditions have a deleterious effect on platelet mitochondrial function and platelet activation.
Platelet donations were collected from healthy donors (n = 5) and stored in gas-permeable collection bags according to American Association of Blood Bank recommendations. Platelet units were sampled from day of collection (day 0) until day 7. High-resolution respirometry was used to assess baseline mitochondrial respiration, maximal oxygen utilization, and individual mitochondrial complex-dependent respiration. Fluorescence-activated cell sorting was performed to analyze mitochondrial content, mitochondrial reactive oxygen species, the expression of P-selectin (both before and after challenge with thrombin receptor-activating peptide), and apoptosis. Data were analyzed using analysis of variance and Pearson correlation (P < 0.05 significant).
Mitochondrial respiration decreased significantly in platelets stored longer than 2 d (P < 0.05). Platelets also demonstrated a persistent decrease in response to stimulation with thrombin receptor-activating peptide by the third day of storage (P < 0.05) as well as an increase in mitochondrial reactive oxygen species and apoptosis (P < 0.05). Mitochondrial respiration significantly correlated with platelet capacity to activate (r = 0.8, P < 0.05).
Platelet mitochondrial respiratory function and activation response decrease significantly in platelets stored for 3 d or more. Because platelet transfusions almost universally occur between the third and fifth day of storage, our findings may have significant clinical importance and warrant further in vivo analysis.
出血性休克是严重创伤后死亡的主要原因,常需要血小板输注以达到止血目的。然而,血小板需要特殊的储存条件,储存时间与血小板质量的损失有关。我们假设标准储存条件对血小板线粒体功能和血小板激活有有害影响。
从健康供体(n=5)中采集血小板捐献物,并根据美国血库协会的建议储存在透气收集袋中。从采集当天(第 0 天)到第 7 天,从血小板单位中取样。使用高分辨率呼吸测定法评估基础线粒体呼吸、最大耗氧量和个体线粒体复合物依赖性呼吸。使用荧光激活细胞分选术分析线粒体含量、线粒体活性氧、P-选择素的表达(在与血栓酶受体激活肽孵育前后)和凋亡。使用方差分析和 Pearson 相关性分析(P<0.05 为显著)进行数据分析。
储存超过 2 天的血小板线粒体呼吸显著降低(P<0.05)。血小板在储存的第三天对血栓酶受体激活肽的刺激也表现出持续的反应性降低(P<0.05),以及线粒体活性氧和凋亡增加(P<0.05)。线粒体呼吸与血小板激活能力显著相关(r=0.8,P<0.05)。
储存 3 天或更长时间的血小板线粒体呼吸功能和激活反应显著降低。由于血小板输注几乎普遍发生在储存的第 3 天至第 5 天之间,因此我们的发现可能具有重要的临床意义,需要进一步的体内分析。