The Reactive Oxygen Species Scavenger N-Acetyl-L-Cysteine Reduces Storage-Dependent Decline in Integrin α IIb β 3-Mediated Platelet Function, Inhibiting Pre-Activation of Integrin and Its β 3 Subunit Cleavage.

2区 生物学 Q1 Biochemistry, Genetics and Molecular Biology
Oxidative Medicine and Cellular Longevity Pub Date : 2025-04-21 eCollection Date: 2025-01-01 DOI:10.1155/omcl/7499648
Ehteramolsadat Hosseini, Zahra Beyranvand, Simone M Schoenwaelder, Fateme Farhid, Mehran Ghasemzadeh
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引用次数: 0

Abstract

Background: Premature activation of integrin α IIb β 3 plays a central role in the induction and development of the platelet storage lesion (PSL) characterized by an exhausted platelet phenotype that affects adhesion and spreading on fibrinogen. Given the role of reactive oxygen species (ROS) in regulating platelet activation per se, we investigated the effects of a ROS scavenger on reducing the functional decline of platelet integrin α IIb β 3 during storage. Methods: Platelet-rich plasma-platelet concentrates (PRP-PCs) were either treated with ROS-reducing agents (1 mM N-acetyl-L-cysteine [NAC] or 30 μM NADPH oxidase [NOX] inhibitor, VAS2870) or kept untreated during storage. CD41/CD61 (total integrin α IIb β 3) expression and PAC-1 binding (specific to active integrin α IIb β 3 conformation) were analyzed by flow cytometry over a 5 day storage period. Molecular changes in integrin β 3 subunit were evaluated by western blotting. Platelet adhesion/spreading to fibrinogen in the presence of ROS inhibitors was also investigated during storage using fluorescence microscopy. Results: A decrease in the molecular weight of integrin β 3 subunit was observed during platelet storage, and was significantly reduced by NAC but not VAS2870, suggesting proteolytic cleavage of β 3 during storage. Further to this, ROS inhibitors decreased integrin activation and increased platelet adhesion to fibrinogen from day 3 of storage, while NAC but not VAS2870 improved platelet spreading. Conclusion: This is the first report of increasing β 3 cleavage of integrin during storage that was inversely correlated with integrin α IIb β 3-mediated platelet function. In this regard, as a generic ROS scavenger, NAC was shown to reduce defects in platelet spreading through inhibition of β 3 cleavage. This is in contrast to VAS2870 which selectively inhibits cytosolic NOX alone, suggesting that the reduced platelet function observed during storage may be due to cumulative effects of mitochondrial ROS. Taken together, these studies suggest that adding NAC to platelets may significantly preserve optimal integrin α IIb β 3 and platelet function during storage. Moreover, as a reversible scavenger, its inhibitory effect can be readily compensated after transfusion.

活性氧清除剂n -乙酰- l-半胱氨酸抑制整合素α IIb β 3介导的血小板功能的储存依赖性下降,抑制整合素的预激活及其β 3亚基切割。
背景:整合素α IIb β 3的过早激活在血小板储存病变(PSL)的诱导和发展中起核心作用,其特征是血小板耗尽表型,影响纤维蛋白原的粘附和扩散。考虑到活性氧(ROS)本身在调节血小板活化中的作用,我们研究了ROS清除剂对血小板整合素α IIb β 3在储存过程中功能下降的影响。方法:将富血小板血浆血小板浓缩物(PRP-PCs)分别用1 mM n -乙酰- l-半胱氨酸(NAC)或30 μM NADPH氧化酶(NOX)抑制剂VAS2870处理或保存。流式细胞术分析5天内CD41/CD61(总整合素α IIb β 3)表达和PAC-1结合(特异性整合素α IIb β 3构象)。western blotting检测整合素β 3亚基的分子变化。在存在ROS抑制剂的情况下,还使用荧光显微镜研究了储存期间血小板对纤维蛋白原的粘附/扩散。结果:血小板储存过程中,整联素β 3亚基分子量降低,NAC显著降低,而VAS2870不显著,提示血小板储存过程中β 3发生了蛋白水解裂解作用。此外,从储存第3天开始,ROS抑制剂降低了整合素的激活,增加了血小板对纤维蛋白原的粘附,而NAC而不是VAS2870改善了血小板的扩散。结论:这是首次报道整合素在储存过程中β 3切割增加,与整合素α IIb β 3介导的血小板功能呈负相关。在这方面,作为一种通用的ROS清除剂,NAC被证明通过抑制β 3切割来减少血小板扩散缺陷。这与VAS2870相反,VAS2870选择性地单独抑制细胞内NOX,这表明在储存期间观察到的血小板功能降低可能是由于线粒体ROS的累积效应。综上所述,这些研究表明,在血小板中添加NAC可以在储存过程中显著保持最佳的整合素α IIb β 3和血小板功能。此外,作为一种可逆的清除剂,其抑制作用在输血后可以很容易地补偿。
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来源期刊
CiteScore
13.20
自引率
0.00%
发文量
1274
审稿时长
3-8 weeks
期刊介绍: Oxidative Medicine and Cellular Longevity is a unique peer-reviewed, Open Access journal that publishes original research and review articles dealing with the cellular and molecular mechanisms of oxidative stress in the nervous system and related organ systems in relation to aging, immune function, vascular biology, metabolism, cellular survival and cellular longevity. Oxidative stress impacts almost all acute and chronic progressive disorders and on a cellular basis is intimately linked to aging, cardiovascular disease, cancer, immune function, metabolism and neurodegeneration. The journal fills a significant void in today’s scientific literature and serves as an international forum for the scientific community worldwide to translate pioneering “bench to bedside” research into clinical strategies.
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