Zbikowska Halina Malgorzata, Nowak Pawel, Wachowicz Barbara
Department of General Biochemistry, University of Lodz, Lodz, Poland.
Blood Coagul Fibrinolysis. 2007 Apr;18(3):271-7. doi: 10.1097/MBC.0b013e3280a69148.
The objective was to study the effects of gamma irradiation, in the presence of sodium ascorbate, on coagulation/fibrinolytic activity of fresh frozen plasma to be applied to inactivate the transfusion-transmitted viruses in plasma-derived products. Plasma was irradiated (50 kGy total dose, on dry ice) using a 60Co source. The plasma proteins were analysed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and western blot and the following parameters estimated: prothrombin time, functional fibrinogen concentration, thrombin-induced fibrinogen polymerization, plasminogen activity, and tissue-type plasminogen activator-induced conversion of plasminogen to plasmin. In irradiated plasma a moderate fragmentation of the most labile plasma proteins was found. The prothrombin time was prolonged (1.5-fold), functional fibrinogen was significantly reduced (60%), fibrinogen polymerization was impaired, plasminogen was predominantly maintained (90%) and tissue-type plasminogen activator-induced conversion of plasminogen to plasmin was unchanged. Ascorbate (25 mmol/l) raised the level of functional fibrinogen in irradiated plasma (to 50%; P=0.0245) and slightly accelerated its polymerization. The small protective effect of ascorbate might be due to inhibition of the radiation-induced fibrinogen oxidation and/or fragmentation but addition of other antioxidants/stabilizers would be crucial when a high irradiation dose, an effective treatment for inactivation of the most resistant viruses, is applied.
目的是研究在抗坏血酸钠存在的情况下,γ射线辐照对新鲜冰冻血浆凝血/纤维蛋白溶解活性的影响,以用于灭活血浆制品中输血传播的病毒。使用60Co源对血浆进行辐照(总剂量50 kGy,在干冰上)。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分析血浆蛋白,并评估以下参数:凝血酶原时间、功能性纤维蛋白原浓度、凝血酶诱导的纤维蛋白原聚合、纤溶酶原活性以及组织型纤溶酶原激活剂诱导的纤溶酶原向纤溶酶的转化。在辐照后的血浆中发现最不稳定的血浆蛋白有中度片段化。凝血酶原时间延长(1.5倍),功能性纤维蛋白原显著降低(60%),纤维蛋白原聚合受损,纤溶酶原主要得以保留(90%),并且组织型纤溶酶原激活剂诱导的纤溶酶原向纤溶酶的转化未发生变化。抗坏血酸(25 mmol/l)提高了辐照后血浆中功能性纤维蛋白原的水平(至50%;P = 0.0245)并略微加速了其聚合。抗坏血酸的这种微小保护作用可能是由于抑制了辐射诱导的纤维蛋白原氧化和/或片段化,但当应用高辐照剂量(一种有效灭活最具抗性病毒的处理方法)时,添加其他抗氧化剂/稳定剂将至关重要。