降低血小板活性氧生成对减少冷藏血小板被吞噬的作用  

The effect of reducing platelet reactive oxygen species on decreasing of the refrigerated platelets being phagocyted

作　　者：谢如锋[1] 王志成[2] 孙洁[1] 杨懿铭[1] 杨洁[1] Xie Rufeng;Wang Zhicheng;Sun Jie;Yang Yiming;Yang Jie(Blood Engineering Laboratory,Shanghai Blood Center,Shanghai 200051;Dept of Transfusion,Huashan Hospital,Fudan University,Shanghai 200040)

机构地区：[1]上海市血液中心血液工程学科,上海200051 [2]复旦大学附属上海华山医院输血科,上海200040

出　　处：《安徽医科大学学报》2025年第1期123-129,共7页Acta Universitatis Medicinalis Anhui

基　　金：上海市卫生健康委员会科研课题计划项目(编号:201940111、202140008);上海市科学技术委员会科研计划项目(编号:15ZR1438300)。

摘　　要：目的探究N-乙酰半胱氨酸(NAC)的添加在血小板冷藏保存期间对活性氧(ROS)异常增加的影响,进而防止血小板被巨噬细胞和肝细胞识别并清除的作用。方法将浓缩血小板分成室温组(22C组)、冷藏组(4C组)和NAC添加组;NAC添加组为冷藏血小板中分别添加5 mmol/L(N5组)和25 mmol/L(N25组)的NAC;各组血小板保存5 d后,流式细胞仪检测血小板ROS、活化等指标。佛波酯诱导的THP-1细胞或原代培养的HepG2细胞用于检测对各组血小板的吞噬作用。结果保存5 d后,4C组血小板ROS水平较22C组升高(P<0.05);加入NAC后,ROS水平则降低(P<0.05)。与4C组相比,N5组血小板CD62P、磷脂酰丝氨酸(PS)的表达以及β-N-乙酰萄糖胺(β-GlcNAc)的暴露程度均降低(P<0.05)。THP-1和HepG2对N5组血小板的吞噬率也低于4C组(P<0.05)。但添加5 mmol/L NAC对冷藏5 d血小板的计数、pH值、CD42b表达、β-半乳糖(β-Gal)暴露和血小板凝血功能无显著影响。结论冷藏血小板添加NAC可显著降低血小板ROS水平,有效减少冷藏血小板被细胞吞噬,提示NAC能防止血小板回输体内后被吞噬清除。Objective To explore the effect of the addition of N-acetylcysteine(NAC)on the abnormal increase of reactive oxygen species(ROS)during platelet cold storage,and to clarify its function of preventing the platelets from being recognized and cleared by macrophages and hepatocytes.Methods Platelets Concentrates were divided into room temperature group(22C),cold storage group(4C)and NAC addition group.In NAC addition group,the platelets were supplemented with 5 mmol/L(N5)and 25 mmol/L(N25)NAC.After 5-day storage,the levels of ROS,activation markers and other index of platelets in each group were detected by flow cytometry.Platelet phagocytosis was detected by PMA-activated THP-1 cells or by primary cultured HepG2 cells.Results After 5 days of storage,ROS increased significantly in 4C group than those in 22C group(P<0.05),and after NAC addition,ROS level reduced significantly(P<0.05).The expression of CD62P and PS and the exposure ofβ-GlcNAc on platelets in N5 group significantly decreased compared with those in 4C group(P<0.05).The platelet phagocytosis by THP-1 and HepG2 cells were also significantly lower in N5 group than those in 4C group(P<0.05).However,the addition of 5 mmol/L NAC did not significantly affect the count,pH,CD42b expression,β-Gal exposure and coagulation function of cold-stored platelet after 5-day storage.Conclusion The addition of NAC to refrigerated platelets can significantly decrease the level of platelet ROS and effectively reduce their phagocytosis by cells,suggesting that NAC addition may protect refrigerated platelets from being cleared by phagocytosis after transfusion.

关 键 词：血小板 冷藏保存 活性氧 N-乙酰半胱氨酸 吞噬 冷藏损伤 

分 类 号：R457.11[医药卫生—治疗学]