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目的:探讨维生素B_2光化学病原体灭活技术(PRT)处理的单采血小板对白细胞和血小板源细胞因子释放的影响。方法:随机选取捐献血小板的献血员20人,各取60 ml去白细胞单采血小板,平均分成2份:一份不做处理作为对照组,另一份采用维生素B_2联合紫外光照射的光化学法处理作为实验组。在血站标准条件下保存7d,分别于1、3、5和7 d取样,分析血小板计数(PC)、血小板分布宽度(PDW)、平均血小板体积(MPV);应用ELISA方法检测白细胞源细胞因子(IL-1β、IL-2、IL-6、IL-8、TNF-α和IFN-γ)和血小板源细胞因子(CCL3、CCL5、TGF-β-1和PF4)含量以及可溶性P-selectin含量;流式细胞术检测血小板膜磷脂酰丝氨酸(PS)表达水平。结果:实验组与对照组相比,PC、PDW和MPV没有统计学差异。随着血小板贮存期的延长,血小板源性细胞因子含量逐渐增加,5-7 d时达到一个新的高度平台,且实验组中血小板源性细胞因子的含量显著高于对照组(P<0.05)。在血小板贮存的3、5和7 d时,实验组中PS表达阳性率显著高于对照组(P=0.0052,P=0.010,P=0.011)。在5和7 d时,实验组中P-selectin含量显著高于对照组(P=0.0138,P=0.0036)。白细胞源性细胞因子含量在实验组和对照组中相比较差异无统计学意义。结论:经PRT处理的去白细胞血小板中,细胞因子的释放主要来源于血小板,在贮存5和7 d时,血小板源性细胞因子的积累达到一个新的平台,其原因最可能是血小板的活化和凋亡。
OBJECTIVE: To investigate the effect of platelets derived from vitamin B_2 photochemical pathogen inactivation (PRT) on leukocyte and platelet-derived cytokine release. Methods: Totally 20 donor blood donors donated platelets, each taking 60 ml leukocyte apheresis platelets, evenly divided into two parts: one without treatment as control group and the other with photochemical treatment with vitamin B 2 combined with UV light As experimental group. Blood samples were stored for 7 days under the standard blood conditions. Samples were taken at 1, 3, 5, and 7 days to analyze platelet count (PC), platelet distribution width (PDW) and mean platelet volume (MPV). ELISA was used to detect leukocyte-derived cytokines The levels of IL-1β, IL-2, IL-6, IL-8, TNF-α and IFN-γ and platelet derived cytokines (CCL3, CCL5, TGF-β-1 and PF4) Flow cytometry was used to detect the level of phosphatidylserine (PS) in platelet membrane. Results: There was no significant difference in PC, PDW and MPV between the experimental group and the control group. With the prolongation of platelet storage, the content of platelet-derived cytokines gradually increased and reached a new height plateau 5-7 d, and the content of platelet-derived cytokines in the experimental group was significantly higher than that in the control group (P <0.05) . At 3, 5 and 7 days after platelet storage, the positive rate of PS expression in the experimental group was significantly higher than that in the control group (P = 0.0052, P = 0.010, P = 0.011). At 5 and 7 days, P-selectin levels in the experimental group were significantly higher than those in the control group (P = 0.0138, P = 0.0036). Leukocyte-derived cytokine content in the experimental group and the control group, the difference was not statistically significant. CONCLUSIONS: The release of cytokines mainly from platelets in PRT-treated leukocyte platelets reaches a new platform for platelet-derived cytokine accumulation at 5 and 7 days of storage, most likely due to platelet activation and Apoptosis.