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储存血小板浓缩物中的补体激活。

Complement activation in stored platelet concentrates.

作者信息

Miletic V D, Popovic O

机构信息

Department of Tissue Typing and Immunochemistry, Blood Transfusion Institute, Belgrade, Yugoslavia.

出版信息

Transfusion. 1993 Feb;33(2):150-4. doi: 10.1046/j.1537-2995.1993.33293158048.x.

Abstract

Activation of platelets during preparation and/or storage of platelet concentrates in plastic containers at room temperature has recently been recognized. Many different biologic causes of this activation have been postulated. Activated complement, as a multi-enzyme system, is one of the possible sources of molecules leading to platelet activation. To detect complement activation, functional complement activity and the generation of complement-derived ligands were investigated in platelet concentrate supernatant plasma during 5 days of storage at room temperature. Hemolytic tests for functional classical and alternative pathway activity were used, as was the kinetic test for complement-mediated inhibition of immune complex precipitation. The presence of C3 activation products (C3, C3c, C3dg) was investigated in plasma by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting procedures and on platelets by immunofluorescence. Activation of complement was evident during storage, and C3c and C3d fragments were clearly demonstrated in plasma. The amount of C3d fragments on platelets gradually rose during the first 3 days of storage. At the end of 5 days of storage, the platelets became C3d negative. There are two possible mechanisms of C3d disappearance--shedding and/or further degradation of C3d fragments. Those results indicated that complement activation and the generation of complement-dependent ligand-receptor interaction may be mechanisms for platelet activation in concentrates stored at room temperature.

摘要

近期已认识到,在室温下于塑料容器中制备和/或储存血小板浓缩物的过程中,血小板会被激活。关于这种激活存在许多不同的生物学原因假说。作为一种多酶系统,活化补体是导致血小板激活的分子的可能来源之一。为检测补体激活情况,在室温下储存5天期间,对血小板浓缩物上清血浆中的功能性补体活性及补体衍生配体的生成进行了研究。采用了针对功能性经典途径和替代途径活性的溶血试验,以及补体介导的免疫复合物沉淀抑制动力学试验。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和蛋白质印迹法在血浆中研究C3激活产物(C3、C3c、C3dg)的存在情况,并通过免疫荧光法在血小板上进行研究。储存期间补体激活明显,血浆中清晰显示出C3c和C3d片段。储存前3天血小板上C3d片段的量逐渐增加。储存5天结束时,血小板变为C3d阴性。C3d消失有两种可能机制——C3d片段的脱落和/或进一步降解。这些结果表明,补体激活及补体依赖性配体-受体相互作用的产生可能是室温下储存的浓缩物中血小板激活的机制。

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