核黄素光化学法处理对全血中淋巴细胞功能的影响

The Effect of Riboflavin Photochemical Method on the Function of Lymphocytes in Whole Blood

目的 研究核黄素光化学法在一定剂量条件下处理全血后对全血中淋巴细胞功能的影响。方法 随机选取多人份当天采集的全血,实验组全血中加入核黄素溶液,终浓度160 μmol/mL,并注入光照血袋中,全血厚度约1.3 mm,放置灭活仪中接受18 J/cm^2的紫外照射剂量;对照组使用相同来源的全血,不加核黄素不进行紫外照射。分离提取全血中淋巴细胞,用CCK-8试剂盒检测淋巴细胞增殖活性,计算其增殖抑制率;用终浓度为1 μg/mL的植物血凝素(PHA)同步刺激两组淋巴细胞后,检测其细胞因子的分泌量。结果 核黄素光化学法处理后,淋巴细胞增殖抑制率为(74.18±14.00)%;未经PHA刺激,淋巴细胞中IFN-γ、IL-8、IL-12、IL-10、IL-13、IL-4细胞因子分泌量分别下降了63%、35%、61%、42%、77%、8%;加PHA刺激后,淋巴细胞中IFN-γ、IL-8、IL-12、IL-10、IL-13、IL-4细胞因子分泌量分别下降了72%、43%、40%、78%、82%、2%;两组IL-4的分泌量无统计学差异。结论 核黄素光化学法在一定的剂量条件下处理全血,会抑制全血中淋巴细胞的增殖功能和细胞因子分泌功能,但并没有完全丧失,仍可保持一定的功能活性,对未来利用核黄素光化学法建立血液体外循环处理回输技术有一定的参考意义。

Objective To study the effect of riboflavin photochemical method on the function of lymphocyte in whole blood treated with a certain dose.Methods In the experimental group,the riboflavin solution was added to whole blood with a final concentration of 160μmol/mL,then whole blood was injected into illumination blood bag.The thickness of whole blood was approximate 1.3 mm and the ultraviolet dose was 18 J/cm^2.The control group came from the same whole blood units without riboflavin and ultraviolet radiation.Lymphocytes in whole blood were separated to detect lymphocyte proliferation activity using CCK-8 kit and calculate proliferation inhibition rate.Stimulated by Phytohemagglutinin(PHA)with a final concentration of 1μg/mL,cytokines secreted from lymphocytes were detected using cytokines kits.Results The lymphocyte proliferation inhibition rate was(74.18±14.00)%after riboflavin photochemical treatment.Cytokine IFN-γ,IL-8,IL-12,IL-10,IL-13 and IL-4decreased by 63%,35%,61%,42%,77%and 8%respectively;After stimulated by PHA,cytokine IFN-γ,IL-8,IL-12,IL-10,IL-13,IL-4 decreased by 72%,43%,40%,78%,82%and 2%respectively;there was no statistical difference in the secretion of IL-4 between two groups.Conclusion Riboflavin photochemical treatment at a certain dose would inhibit partly the proliferation and cytokine secretion of lymphocytes in the whole blood.