Molecular Medicine Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland 20892, USA.
Transfusion. 2013 Mar;53(3):637-44. doi: 10.1111/j.1537-2995.2012.03777.x. Epub 2012 Jul 15.
Nitric oxide (NO), a potent signaling molecule, is known to inhibit platelet (PLT) function in vivo. We investigated how the levels of NO and its metabolites change during routine PLT storage. We also tested whether the material of PLT storage containers affects nitrite content since many plastic materials are known to contain and release nitrite.
For nitrite and nitrate measurement, leukoreduced apheresis PLTs and concurrent plasma (CP) were collected from healthy donors using a cell separator. Sixty-milliliter aliquots of PLT or CP were stored in CLX or PL120 Teflon containers at 20 to 24°C with agitation and daily samples were processed to yield PLT pellet and supernatant. In a separate experiment, PLTs were stored in PL120 Teflon to measure NO generation using electron paramagnetic resonance (EPR).
Nitrite level increased markedly in both PLT supernatant and CP stored in CLX containers at a rate of 58 and 31 nmol/L/day, respectively. However, there was a decrease in nitrite level in PLTs stored in PL120 Teflon containers. Nitrite was found to leach from CLX containers and this appears to compensate for nitrite consumption in these preparations. Nitrate level did not significantly change during storage.
PLTs stored at 20 to 24°C maintain measurable levels of nitrite and nitrate. The nitrite decline in nonleachable Teflon containers in contrast to increases in CLX containers that leach nitrite suggests that it is consumed by PLTs, residual white blood cells, or red blood cells. These results suggest NO-related metabolic changes occur in PLT units during storage.
一氧化氮(NO)作为一种强效的信号分子,在体内被认为能够抑制血小板(PLT)的功能。我们研究了在常规 PLT 储存过程中 NO 及其代谢物的水平如何变化。我们还测试了 PLT 储存容器的材料是否会影响亚硝酸盐含量,因为许多塑料材料已知会含有并释放亚硝酸盐。
为了测量亚硝酸盐和硝酸盐,使用细胞分离器从健康供体中采集白细胞减少的单采 PLT 和同时采集的血浆(CP)。将 60 毫升 PLT 或 CP 等分试样分别储存在 CLX 或 PL120 聚四氟乙烯容器中,在 20 至 24°C 下搅拌,并每天处理样品以获得 PLT 沉淀和上清液。在另一个实验中,将 PLT 储存在 PL120 聚四氟乙烯中,使用电子顺磁共振(EPR)测量 NO 的生成。
在 CLX 容器中储存的 PLT 上清液和 CP 中,亚硝酸盐水平分别以 58 和 31 nmol/L/天的速度显著增加。然而,在 PL120 聚四氟乙烯容器中储存的 PLT 中亚硝酸盐水平下降。在 CLX 容器中发现亚硝酸盐浸出,这似乎补偿了这些制剂中亚硝酸盐的消耗。在储存过程中,硝酸盐水平没有显著变化。
在 20 至 24°C 下储存的 PLT 保持可测量水平的亚硝酸盐和硝酸盐。不可浸出的聚四氟乙烯容器中亚硝酸盐的下降与浸出亚硝酸盐的 CLX 容器中的增加形成对比,这表明 PLT、残留的白细胞或红细胞消耗了亚硝酸盐。这些结果表明,在储存过程中,PLT 单位中发生了与 NO 相关的代谢变化。