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*

doi:10.3969/j.issn.1671-2587.2024.05.007

临床输血与检验 ›› 2024, Vol. 26 ›› Issue (5): 617-623.DOI: 10.3969/j.issn.1671-2587.2024.05.007

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^*

谢如锋, 张景怡, 杨懿铭, 高跞, 蒋雪玉, 孙娟, 杨洁

上海市血液中心,输血研究所,血液工程学科,上海 200051

收稿日期:2024-06-07 出版日期:2024-10-20 发布日期:2024-09-20
通讯作者: 杨洁,主要从事细胞治疗、血液保存等输血医学研究,（E-mail）yangjie@sbc.org.cn。
作者简介:谢如锋,主要从事血液保存等输血医学研究,（E-mail）xierufeng@sbc.org.cn。张景怡,主要从事临床输血检验技术工作,（E-mail）zhangjingyi@sbc.org.cn。
基金资助:
*本课题受上海市卫生健康委员会面上项目（No.201940111、202140008）资助

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

XIE Rufeng, ZHANG Jingyi, YANG Yiming, GAO Li, JIANG Xueyu, SUN Juan, YANG Jie

Blood Engineering Laboratory, Shanghai Blood Center, Shanghai 200051

Received:2024-06-07 Online:2024-10-20 Published:2024-09-20

摘要/Abstract

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,3 960±259.6 vs. 5 846±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（5 846±757.4 vs. 3 945±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。

关键词: 近红外低能量激光, 冷藏血小板, 冷藏损伤, 活性氧

Abstract: 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). PMA-induced 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, 3 960±259.6 vs. 5 846±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 (5 846±757.4 vs. 3 945±459.4, P<0.01) and diminished the exposure of CD62P, phosphatidylserine (PS) expression and GPIb/ɑ glycoprotein 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/ɑ desialylation, thereby reducing cold storage-induced lesions and preventing phagocytosis.

Key words: Near-infrared low level laser, Cold-stored platelet, Cold storage-induced lesions, Reactive oxygen species

中图分类号:

R457

谢如锋, 张景怡, 杨懿铭, 高跞, 蒋雪玉, 孙娟, 杨洁. 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^*[J]. 临床输血与检验, 2024, 26(5): 617-623.

XIE Rufeng, ZHANG Jingyi, YANG Yiming, GAO Li, JIANG Xueyu, SUN Juan, YANG Jie. 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[J]. JOURNAL OF CLINICAL TRANSFUSION AND LABORATORY MEDICINE, 2024, 26(5): 617-623.

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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

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