Loss of 12-Lipoxygenase Improves the Post-Transfusion Function of Stored Platelets.
Loss of 12-Lipoxygenase Improves the Post-Transfusion Function of Stored Platelets.
复制标题
12-脂氧合酶的损失可改善储存血小板的输血后功能。
DOI：
10.1161/atvbaha.123.319021
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发表时间：
2023
期刊：
影响因子：
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通讯作者：
Stolla,Moritz
中科院分区：
文献类型：
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作者：
Larsen,HannahJ;Byrne,Daire;Özpolat,Tahsin;Chauhan,Aastha;Bailey,SLawrence;Rhoads,Nicole;Reed,Franklin;Stolla,MassielC;Adili,Reheman;Holinstat,Michael;Fu,Xiaoyun;Stolla,Moritz
输血用血小板储存5至7天。先前的研究表明，HETE水平的存储袋中的血小板在体内的性能呈负相关，表明生物活性脂质介质的失调可能有助于存储病变。在目前的研究中，我们试图了解12-LOX的基因缺失和药理学抑制是如何（12-脂氧合酶）在储存期间和输血后影响血小板。方法来自12-LOX +/+的血小板（野生型[WT]）和12-LOX −/−小鼠储存24和48小时，并使用液相色谱-串联质谱-多反应监测进行分析，或输注到血小板减少的hIL 4R中。（人白细胞介素4受体）-转基因小鼠。通过流式细胞术和体内血栓形成和止血模型评估血小板功能。为了测试考克斯-1（环氧合酶-1）途径的作用，用乙酰水杨酸处理供体小鼠。用12-LOX抑制剂VLX-1005或载体处理人血小板，储存，并输注给NOD/SCID（非肥胖糖尿病/严重联合免疫缺陷）小鼠。在来自12-LOX −/−小鼠的储存血小板中，多不饱和脂肪酸显著增加，而氧化脂质浓度在WT血小板中显著更高。输注给血小板减少小鼠后，我们观察到12-LOX −/−血小板中α IIb β 3整合素的基线活化显著高于WT血小板。来自12-LOX −/−小鼠的储存的血小板比储存的WT血小板显著更快地阻塞血管。在止血模型中，静脉损伤部位储存的12-LOX −/−血小板显著多于WT血小板，导致失血减少。抑制考克斯-1可以消除储存的12-LOX −/−血小板中整合素活化和血栓烷生成的增加，突出了该途径对改善输血后功能的关键作用。与我们的小鼠研究一致，与VLX-1005储存的人血小板，显示出增加的整合素活化与车辆处理的血小板相比transmiss.CONCLUSIONSDeleting 12-LOX改善输血后功能的储存鼠血小板通过增加血栓素生成通过考克斯-1依赖性花生四烯酸代谢。未来的研究应该确定12-LOX抑制血小板输注给人类的可行性和安全性。
BACKGROUNDPlatelets for transfusion are stored for 5 to 7 days. Previous studies have shown that HETE levels in the storage bag negatively correlate with platelet performance in vivo, suggesting that the dysregulation of bioactive lipid mediators may contribute to the storage lesion. In the current study, we sought to understand how genetic deletion and pharmacological inhibition of 12-LOX (12-lipoxygenase) affects platelets during storage and after transfusion.METHODSPlatelets from 12-LOX+/+(wild-type [WT]) and 12-LOX−/−mice were stored for 24 and 48 hours and profiled using liquid chromatography-tandem mass spectrometry–multiple reaction monitoring or transfused into thrombocytopenic hIL4R (human interleukin 4 receptor)-transgenic mice. Platelet function was assessed by flow cytometry and in vivo thrombosis and hemostasis models. To test the role of the COX-1 (cyclooxygenase-1) pathway, donor mice were treated with acetylsalicylic acid. Human platelets were treated with the 12-LOX inhibitor, VLX-1005, or vehicle, stored, and transfused to NOD/SCID (nonobese diabetic/severe combined immunodeficiency) mice.RESULTSPolyunsaturated fatty acids increased significantly in stored platelets from 12-LOX−/−mice, whereas oxylipin concentrations were significantly higher in WT platelets. After transfusion to thrombocytopenic mice, we observed significantly more baseline αIIbβ3 integrin activation in 12-LOX−/−platelets than in WT platelets. Stored platelets from 12-LOX−/−mice occluded vessels significantly faster than stored WT platelets. In hemostasis models, significantly more stored 12-LOX−/−than WT platelets accumulated at the site of venous injury leading to reduced blood loss. Inhibition of COX-1 abrogated both increased integrin activation and thromboxane generation in stored 12-LOX−/−platelets, highlighting the critical role of this pathway for improved post-transfusion function. Consistent with our mouse studies, human platelets stored with VLX-1005, showed increased integrin activation compared with vehicle-treated platelets after transfusion.CONCLUSIONSDeleting 12-LOX improves the post-transfusion function of stored murine platelets by increasing thromboxane generation through COX-1–dependent arachidonic acid metabolism. Future studies should determine the feasibility and safety of 12-LOX–inhibited platelets transfused to humans.