亚甲蓝光化学法灭活病毒的研究——抗坏血酸对血浆凝血因子的保护作用

Study on inactivating by methylene blue photochemical methods──The protective effect of ascorbic acid on coagulation factors of plasma

目的 探讨抗坏血酸对亚甲蓝光化学法处理血浆时指示病毒灭活的效果 ,以及对部分凝血因子的保护作用。方法 采用CPE法评价病毒的灭活效果 ,用Clauss法测定纤维蛋白原的含量和一期法判定FⅧ :C的凝血活性。结果 在单采人血浆中加入 1μmol·L-1亚甲蓝和 2 4 0 μmol·L-1抗坏血酸 ,再给以 4 0 0 0 0lx荧光强度照射 6 0min ,VSV滴度下降 >8lgTCID50 ·ml-1,并且纤维蛋白原和FⅧ :C的回收率分别为 83.5 5 %和 81.6 7% ,比常规亚甲蓝 /荧光光化学法显著提高 (P <0 .0 1) ,而且凝血酶原时间延长值在正常允许值范围 ,活化部分凝血酶时间延长值也接近正常允许值范围。加入抗坏血酸和色氨酸可灭活病毒 ,而甘露醇不能。结论 在亚甲蓝光化学法中 ,抗坏血酸不影响病毒的灭活 ,而且对部分凝血因子有保护作用。因此 ,抗坏血酸可以有效提高单份冷冻新鲜人血浆的质量。

OBJECTIVE To determine whether ascrobate could influence the inactive efficacy of the indicating viruses by methylene blue(MB)-photochemical method,and whether could protect partial coagulation factors against photooxidation.METHODS Cytopathic effect was used to test the effect of viruses inactivation.Fibrinogen concentration was measured with the Clauss method and factor Ⅷ clotting activity was observed by the one-stage method.RESULTS The results showed that,with the addition of 1 μmol·L -1MB and 240 μmol·L -1Vit C solution into human single plasma unit followed by irradiated at an intensity of 40000 lx for 60 min,the titer of VSV could be decreased by more than 8 lgTICD 50·ml -1.The recovery rates of fibrinogen and FⅧ:C were 83.55% and 81.67%,respectively,which were significantly higher than that of MB/fluorescent light treatment (P<0.01).Prothrombin time (PT) was prolonged within the normal values and activated partial thromboplastin time (APTT) was prolonged slightly beyond the normal values.The protective efficacy of mannitol was not so good as ascrobate and tryptophan influenced the effect of inactivating virus.CONCLUSION These findings indicated that ascrobate protected partial coagulation factors of human plasma from MB-photosensitized damage without affecting virus inactivation.Ascorbic acid would be beneficial for improving the quality of plasma subjected to photodynamic treatment.