首页> 中文期刊>安徽医科大学学报 >核黄素联合紫外光灭活血小板悬液中的病毒及抑制细胞因子的实验研究

核黄素联合紫外光灭活血小板悬液中的病毒及抑制细胞因子的实验研究

     

摘要

Objective To research the safety and efficacy of riboflavin ultraviolet light joint inactivation of virus, and the inhibition of leukocytes derived cytokines in platelets preservation and transfusion and to observe the bio-chemical alterations of the platelets following the inactivation measures. Methods In experimental group human cytomegalovirus ( HCMV AD169 ) was injected into the platelets suspension in final concentration of 150μmol/L of riboflavin solution. After mixing, the sample was put into the apheresis platelets, followed by ultraviolet light steri-lization, and the viral titer was tested in platelet suspension. The production cytokines were detected by ELISA on 0, 3 and 5 d after irradiation, and the change of platelets parameter was observed. Negative control group was com-posed of fresh apheresis platelets from the same collection without any treatments, cytokines were synchronously de-tected. Two groups of platelets were subjected to phytohemagglutinin ( PHA) for synchronous stimulation. Results A dose of 1 500 mJ/cm2 ultraviolet light combined with riboflavin gave rise to a effective sterilization of virus in the platelet suspension. The expression of leukocyte-derived cytokines was noted slight increase in the control group with the extension of saving time post-treatment. In the experimental group No. 3, 5 d the cytokine content had no significant difference relative to (0 d) before saving, while at the same retention time the cytokines content in the control group was higher than that in the experimental group ( P<0.05 );the two groups of platelet suspension after PHA synchronous stimulation:compared to the control group without PHA stimulated, the level of cytokines was in-creased significantly (P<0.05);while with respect to the experimental group without PHA stimulated, the level of cytokines showed no significant change. Conclusion Riboflavin plus ultraviolet light treatment can significantly in-activate selected virus, inhibit the production residual leukocytes of PLT release cytokines during storage, and in vitro apheresis platelets' various parameters compared with negative controls show no significant difference.%目的研究核黄素联合紫外光进行血小板病毒灭活的安全性、有效性及对血小板保存中白细胞释放细胞因子的抑制作用;观察灭活后血小板体外各参数的变化。方法实验组:将人巨细胞病毒标准株( HCMV AD169)注入核黄素溶液,混匀后加入到单采血小板中,以一定辐照剂量的紫外光照射,检测照射前后病毒滴度和照射后不同时间细胞因子的变化,并观察体外血小板部分参数的变化。对照组:为相同来源新鲜单采血小板,同步检测其细胞因子的含量和血小板体外各参数。以植物凝集素( PHA)同步刺激两组血小板,用ELISA法检测细胞因子的含量。结果实验组经150μmol/L的核黄素结合辐照剂量为1500 mJ/cm2的紫外光(250<λ<350 nm)照射10 min 可有效灭活血小板中病毒;对照组细胞因子含量随着保存时间的延长而显著增加;实验组第3天和第5天的细胞因子含量相对于保存前(0 d)差异无统计学意义;而在同一保存时间对照组中细胞因子含量高于实验组(P<0.05);实验组和对照组血小板悬液经过PHA同步刺激后:接受PHA刺激的对照组与未接受PHA刺激的对照组相比,细胞因子含量显著增加( P <0.05);而接受PHA刺激的实验组与未接受PHA刺激的实验组相比,细胞因子含量无显著变化。结论核黄素结合紫外光照射可以有效灭活血小板中病毒,抑制血小板保存中白细胞释放细胞因子的能力,而单采血小板体外诸参数和阴性对照比较无明显差异。

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