Protein composition and functional parameters of RBC membranes in liver and kidney transplantation

A. Deryugina, O. P. Abaeva, S. V. Romanov, M. Vedunova, E. Ryabova, S. Vasenin, N. A. Titova
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Abstract

Organ transplantation is an effective treatment for many end-stage diseases. However, reperfusion injury constitutes a major complication of transplantation, which is associated with microcirculatory disorders and aggregation of blood corpuscles. Red blood cells (RBC) play an essential role in maintaining hemodynamic and rheological properties of the blood. Moreover, the study of mechanisms of changes in RBC functional indices is an urgent task. The main indicator of RBC functioning is the stability of RBC membrane structure. The issue of RBC membrane modification in organ transplantation has not been studied so far. Objective: to study the protein composition of RBC membranes, their aggregation and electrokinetic parameters in liver and kidney recipients, as well as in related kidney and liver fragment donors before and after operation. Research materials. Blood of 12 kidney recipients and 5 related kidney donors, 8 liver recipients and 4 related liver fragment donors – 1–2 hours before surgery, 1 week, 1, 2, 7, 10, 12 months after surgery. The control group consisted of 8 healthy volunteers. Research methods. Protein separation was done by Laemmli electrophoresis. RBC electrophoretic mobility, which characterizes the electrokinetic properties of cells, was measured by microelectrophoresis. Aggregation was calculated microscopically by counting unaggregated RBCs. Obtained values were compared by Mann-Whitney U test. Results. Examination of the RBC membrane of kidney recipients revealed a significant decrease in the amount of Band 3 protein and glycophorin before and after transplantation. Band 3 protein levels reduced at 1 month, glycophorin reduced at 7 months after surgery, with a maximum decrease in these protein fractions by more than 50% by 7 days compared with control values. There was also a decrease in spectrin content for 2 months after surgery with a maximum decrease of 30% by 1 month. In liver recipients, analysis of RBC membrane proteins revealed a decrease in the amount of glycophorin before surgery and further decrease at 2 months of post-transplant period. The maximum decrease in this index was 72% by 7 days after surgery. In addition, there was a fall in spectrin and Band 3 protein levels at 1 month by more than 60% relative to the control values. In donors, there were changes in the protein fraction of RBC membranes in the long-term post-operative period: spectrin and Band 3 protein levels reduced by 2 times at month 2 in kidney donors, while glycophorin levels reduced by 2.3 times at month 1 after operation in liver donors. Similarly, both groups of donors had increased actin levels at month 1 after surgery. The revealed changes in protein levels in the protein phase of RBC membranes were combined with functional indices of RBCs. In kidney recipients, decreased RBC electrophoretic mobility and increased aggregation were detected at 2 months. In liver recipients, the changes in these indicators were at 1 month. A decrease in RBC electrophoretic mobility was detected in donors of both groups. Conclusion. Changes in RBC membrane electronegativity are associated with changes in glycophorin and Band 3 protein levels, whereas in RBC aggregation process in liver/kidney recipients, the structural and functional disorders in the interrelationships of such membrane proteins as spectrin, Band 3 protein, and glycophorin, are significant factors. Alteration of actin determines inhibition of RBC aggregation growth in donors.
肝、肾移植中红细胞膜的蛋白质组成和功能参数
器官移植是许多终末期疾病的有效治疗方法。然而,再灌注损伤是移植的主要并发症,它与微循环障碍和血球聚集有关。红细胞(RBC)在维持血液的血液动力学和流变学特性中起着重要的作用。因此,研究红细胞功能指标变化的机制是一个迫切的任务。红细胞功能的主要指标是红细胞膜结构的稳定性。红细胞膜修饰在器官移植中的应用目前尚未见研究。目的:研究肝、肾受者及相关肾、肝碎片供者手术前后红细胞膜蛋白组成、聚集及电动力学参数。研究材料。术前1 ~ 2小时、术后1周、1、2、7、10、12个月肾受体受者12例及相关肾供者5例、肝受体受者8例及相关肝碎片供者4例。对照组为8名健康志愿者。研究方法。Laemmli电泳法分离蛋白质。红细胞的电泳迁移率,表征细胞的电动特性,是测量微电泳。通过计数未聚集的红细胞在显微镜下计算聚集。所得值采用Mann-Whitney U检验进行比较。结果。肾受体红细胞膜检查显示移植前后Band 3蛋白和糖蛋白含量明显降低。术后1个月Band 3蛋白水平下降,术后7个月糖蛋白水平下降,与对照组相比,这些蛋白部分在7天内最大下降幅度超过50%。术后2个月spectrin含量也有所下降,1个月最多下降30%。在肝受体中,红细胞膜蛋白分析显示术前糖蛋白含量下降,移植后2个月进一步下降。术后7天该指标最大降幅为72%。此外,与对照组相比,1个月时spectrin和Band 3蛋白水平下降了60%以上。在供者中,术后长期红细胞膜蛋白部分发生变化:肾供者术后2个月spectrin和Band 3蛋白水平下降2倍,肝供者术后1个月糖蛋白水平下降2.3倍。同样,两组供体在术后1个月时肌动蛋白水平均有所升高。红细胞膜蛋白期蛋白水平的变化与红细胞功能指标相结合。在肾受体中,2个月时检测到红细胞电泳迁移率下降和聚集增加。在肝受体中,这些指标在1个月时发生变化。两组供体红细胞电泳迁移率均下降。结论。红细胞膜电负性的变化与糖蛋白和带3蛋白水平的变化有关,而在肝/肾受体红细胞聚集过程中,谱蛋白、带3蛋白和糖蛋白等膜蛋白相互关系的结构和功能紊乱是重要因素。肌动蛋白的改变决定了供体红细胞聚集生长的抑制。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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