近红外低能量激光调节血小板活性氧水平对防止冷藏损伤的研究  

Enhancing the Prevention of Cold Storage-induced Lesions:An Experimental Investigation into Modulation of Platelet Reactive Oxygen Species via Low-level Near-infrared Laser Irradiation

作　　者：谢如锋[1] 张景怡[1] 杨懿铭[1] 高跞[1] 蒋雪玉[1] 孙娟[1] 杨洁[1] XIE Rufeng;ZHANG Jingyi;YANG Yiming;GAO Li;JIANG Xueyu;SUN Juan;YANG Jie(Blood Engineering Laboratory,Shanghai Blood Center,Shanghai 200051)

机构地区：[1]上海市血液中心,输血研究所,血液工程学科,上海200051

出　　处：《临床输血与检验》2024年第5期617-623,共7页Journal of Clinical Transfusion and Laboratory Medicine

基　　金：上海市卫生健康委员会面上项目(No.201940111、202140008)资助。

摘　　要：目的探究近红外低能量激光(Low Level Laser,LLL)照射处理对冷藏血小板保存期间活性氧的调节作用,进而对减少被巨噬细胞和肝细胞识别吞噬的影响。方法将白膜来源的浓缩血小板分为室温保存组(RTP)、冷藏保存组(CSP)和LLL照射组(CSP-L)。LLL照射组为血小板冷藏12 h后,采用1 J、5 J、10 J不同辐照强度的LLL单次照射并继续冷藏保存。各组保存5 d后,流式细胞仪检测血小板ROS生成、活化指标表达、糖基蛋白及其残基的暴露情况并进行比较。佛波酯诱导的THP-1细胞和HepG2肝细胞用于检测对各组血小板的吞噬作用。结果保存5 d后,CSP组血小板胞内ROS(cyto-ROS)水平较RTP组显著升高(RTP vs.CSP,3960±259.6 vs.5846±757.4,P<0.01),而线粒体ROS(mito-ROS)水平则低于RTP组(RTP vs.CSP,595±47.1 vs.416±50.2,P<0.05)。经低强度LLL(1 J)照射后,CSP-L1组血小板较CSP组显著降低了cyto-ROS水平(5846±757.4 vs.3945±459.4,P<0.01),CD62P、PS表达和GPⅠb/ɑ糖蛋白残基的暴露程度均显著减少(P<0.05),且与RTP组均无显著性差异。THP-1和HepG2对CSP-L1组血小板的吞噬率也显著低于CSP组(P<0.05)。同时,LLL照射对冷藏血小板的计数、pH值和MPV均无明显影响。结论低强度LLL照射处理可显著降低冷藏血小板胞内ROS水平并维持线粒体ROS的低水平生成,并能有效的抑制血小板活化和GPⅠb/ɑ去唾液酸化,减轻冷藏损伤,进而减少冷藏血小板被巨噬细胞和肝细胞识别吞噬。Objective To explore the modulating effects of near-infrared low level laser(LLL)irradiation on reactive oxygen species(ROS)levels in platelets during cold storage,ultimately reducing cold storage-induced lesions and protecting the platelets from phagocytosis by macrophages and hepatocytes.Methods Buffer-coat-derived platelets were divided into three groups:room-temperature-stored platelets(RTP),cold-stored platelets(CSP),and CSP subjected to LLL irradiation(CSP-L).In the CSP-L group,platelets were irradiated with varying intensities of LLL irradiation(1 J,5 J,10 J)after 12 hours of cold-storage and then maintained under cold condition.After 5-day storage,ROS generation,expression of activation markers,exposure of glycosyl proteins and their residues in platelets from each group were analyzed and compared by flow cytometry(FACS).PMAinduced THP-1 cells and HepG2 hepatocytes were used to examine the phagocytosis of platelets in each group.Results After 5 days of storage,the platelet cytoplasmic ROS(cyto-ROS)level increased significantly in the CSP group compared with the RTP group(RTP vs.CSP,3960±259.6 vs.5846±757.4,P<0.01),whereas the mitochondrial ROS(mito-ROS)level was lower than that in the RTP group(RTP vs.CSP,595±47.1 vs.416±50.2,P<0.05).Notably,low-intensity LLL(1 J)irradiation in the CSP-L1 group significantly reduced cyto-ROS levels(5846±757.4 vs.3945±459.4,P<0.01)and diminished the exposure of CD62P,phosphatidylserine(PS)expression and GPIb/ɑglycoprotein residues compared with the CSP group(P<0.05),aligning closely with the RTP group.Furthermore,the phagocytic rate of platelets by THP-1 and HepG2 cells was also obviously lower in the CSP-L1 group than in the CSP group(P<0.05).Importantly,LLL irradiation had no significant effect on the platelet count,pH,or mean platelet volume(MPV)in cold storage.Conclusion Low-intensity LLL irradiation treatment could effectively lower the cyto-ROS levels of cold-stored platelets while maintaining low mito-ROS generation,and inhibit platelet activation and GPⅠb/

关 键 词：近红外低能量激光 冷藏血小板 冷藏损伤 活性氧 

分 类 号：R457[医药卫生—治疗学]