Wu Tiange, Wang Xiaoning, Ren Kai, Huang Xiaochen, Liu Jiankai
Blood Transfusion Department, The Second Hospital of Jilin University, Changchun, China.
Blood Transfusion Department, The First Hospital of Jilin University, Changchun, China.
Transfus Med Hemother. 2021 Mar 23;48(5):298-305. doi: 10.1159/000515119. eCollection 2021 Oct.
The aim of this study was to investigate the modified proteins in methylene blue/light-treated frozen plasma (MB-FP) compared with fresh frozen plasma (FFP) in order to gain a better application of MB/light-treated plasma in clinic transfusion.
MB-FP and FFP were collected from Changchun central blood station, and a trichloroacetic acid/acetone precipitation method was used to remove albumin for the enrichment of lower abundance proteins. The plasma protein in MB-FP and FFP were separated using two-dimensional gel electrophoresis (2-DE) and the differentially expressed protein spots were analyzed using mass spectrometry. Finally, the differentially expressed proteins were tested using Western blot and enzyme-linked immunosorbent assay (ELISA).
Approximately 14 differentially expressed protein spots were detected in the MB-FP, and FFP was chosen as the control. After 2-DE comparison analysis and mass spectrometry, 8 significantly differentially expressed protein spots were identified, corresponding to 6 different proteins, including complement C1r subcomponent (C1R), inter-alpha-trypsin inhibitor heavy chain H4 (ITI-H4), keratin, type II cytoskeletal 1 (KRT1), hemopexin (HPX), fibrinogen gamma chain (FGG), and transthyretin (TTR). Western blot showed no significant difference in the expression level of KRT1 between MB-FP and FFP ( > 0.05). Both Western blot and ELISA indicated that the level of HPX was significantly higher in FFP than in MB-FP ( < 0.05).
This comparative proteomics study revealed that some significantly modified proteins occur in MB-FP, such as C1R, ITI-H4, KRT1, HPX, FGG, and TTR. Our findings provide more theoretical data for using MB-FP in transfusion medicine. However, the relevance of the data for the transfusion of methylene blue/light-treated plasma remains unclear. The exact modification of these proteins and the effects of these modified proteins on their functions and their effects in clinical plasma infusion need to be further studied.
本研究旨在探究亚甲蓝/光照处理的冰冻血浆（MB-FP）与新鲜冰冻血浆（FFP）相比的修饰蛋白，以便更好地将MB/光照处理的血浆应用于临床输血。
从长春中心血站采集MB-FP和FFP，采用三氯乙酸/丙酮沉淀法去除白蛋白以富集低丰度蛋白。使用二维凝胶电泳（2-DE）分离MB-FP和FFP中的血浆蛋白，并通过质谱分析差异表达的蛋白斑点。最后，使用蛋白质印迹法和酶联免疫吸附测定（ELISA）对差异表达的蛋白进行检测。
在MB-FP中检测到约14个差异表达的蛋白斑点，以FFP作为对照。经过2-DE比较分析和质谱分析，鉴定出8个显著差异表达的蛋白斑点，对应6种不同的蛋白质，包括补体C1r亚成分（C1R）、间α胰蛋白酶抑制剂重链H4（ITI-H4）、角蛋白Ⅱ型细胞骨架1（KRT1）、血红素结合蛋白（HPX）、纤维蛋白原γ链（FGG）和转甲状腺素蛋白（TTR）。蛋白质印迹法显示MB-FP和FFP中KRT1的表达水平无显著差异（>0.05）。蛋白质印迹法和ELISA均表明FFP中HPX的水平显著高于MB-FP（<0.05）。
这项比较蛋白质组学研究表明，MB-FP中存在一些显著修饰的蛋白质，如C1R、ITI-H4、KRT1、HPX、FGG和TTR。我们的研究结果为在输血医学中使用MB-FP提供了更多的理论数据。然而，这些数据与亚甲蓝/光照处理血浆输血的相关性仍不明确。这些蛋白质的确切修饰以及这些修饰蛋白对其功能的影响及其在临床血浆输注中的作用需要进一步研究。
