GSK669, a NOD2 receptor antagonist, inhibits thrombosis and oxidative stress via targeting platelet GPVI.
GSK669, a NOD2 receptor antagonist, inhibits thrombosis and oxidative stress via targeting platelet GPVI.
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DOI：
10.1016/j.bcp.2020.114315
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发表时间：
2020-11
影响因子：
5.8
通讯作者：
Guanxing Pan;Lin Chang;Jianjun Zhang;Yangyang Liu;Liang Hu;Shenmin Zhang;Jian Zhang;J. Qiao-
中科院分区：
文献类型：
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作者：
Guanxing Pan;Lin Chang;Jianjun Zhang;Yangyang Liu;Liang Hu;Shenmin Zhang;Jian Zhang;J. Qiao-
背景和目的我们发现核苷酸结合寡聚化结构域2（NOD 2）的激活可增强血小板的活化。本实验研究了NOD 2拮抗剂GSK 669的抗血小板和抗血栓作用，观察了GSK 669对血小板功能、活性氧（reactive oxygen species，ROS）和促炎细胞因子（pro-inflammatory cytokines，TNF-α）的影响。NOD 2-/-血小板用于确认GSK 669靶标。利用表面等离子体共振（SPR）光谱检测GSK 669与糖蛋白VI（GPVI）之间的相互作用。通过Western印迹检测GPVI下游信号传导。采用小鼠肠系膜小动脉血栓形成模型和肺栓塞模型研究GSK 669的抗血栓形成和抗氧化作用。KeyResultsGSK 669显著抑制胞壁酰二肽诱导的血小板促炎细胞因子释放、血小板聚集、ATP释放和胶原和胶原相关肽（CRP）诱导的ROS生成。血小板扩散和凝块收缩也受到抑制。GSK 669还降低胶原诱导的Src、Syk、PLCγ2和Akt的磷酸化。GSK 669的抗血小板作用是NOD 2非依赖性的，并由GPVI拮抗作用介导。与其作为GPVI拮抗剂的抗血小板活性一致，GSK 669在流动条件下抑制血小板粘附在胶原上。值得注意的是，GSK 669抑制小鼠肠系膜小动脉血栓形成类似于阿司匹林而不出血。在肺栓塞模型中，GSK 669的抗血栓作用得到进一步证实;降低小鼠血浆中丙二醛（MDA）和升高超氧化物歧化酶（SOD）水平显示GSK 669具有显著的抗氧化作用。结论与意义GSK 669除了作为NOD 2拮抗剂的抗炎作用外，还具有靶向GPVI的抗氧化作用，是一种安全有效的抗血小板药物。具有抗氧化和抗炎活性且无出血风险的抗血小板药物可能比目前的抗血小板药物对动脉粥样硬化血栓形成具有治疗优势。
Background and PurposePreviously, we discovered that the activation of nucleotide-binding oligomerization domain 2 (NOD2) enhances platelet activation. We here investigated the antiplatelet and antithrombotic potential of GSK669, a NOD2 antagonist.Experimental ApproachEffects of GSK669 on platelet functions, reactive oxygen species (ROS) and proinflammatory cytokine generation were detected. NOD2-/- platelets were used to confirm GSK669 target. The interaction between GSK669 and glycoprotein VI (GPVI) was detected using surface plasmon resonance (SPR) spectroscopy. GPVI downstream signaling was examined by Western blot. The antithrombotic and antioxidative effects were investigated using mouse mesenteric arteriole thrombosis model and pulmonary embolism model.Key ResultsGSK669 significantly inhibits platelet proinflammatory cytokine release induced by muramyl dipeptide, platelet aggregation, ATP release, and ROS generation induced by collagen and collagen related peptide (CRP). Platelet spreading and clot retraction are also inhibited. GSK669 also decreases collagen-induced phosphorylation of Src, Syk, PLCγ2, and Akt. The antiplatelet effect of GSK669 is NOD2-independent and mediated by GPVI antagonism. Consistent with its antiplatelet activity as a GPVI antagonist, GSK669 inhibits platelet adhesion on collagen in flow condition. Notably, GSK669 inhibits mouse mesenteric arteriole thrombosis similarly to aspirin without bleeding. The antithrombotic effect of GSK669 is further confirmed in the pulmonary embolism model; decreased malonaldehyde (MDA) and increased superoxide dismutase (SOD) levels in mouse plasma reveal a significant antioxidant effect of GSK669.Conclusion and ImplicationsBeyond its anti-inflammatory effect as a NOD2 antagonist, GSK669 is also an efficient and safe antiplatelet agent combined with antioxidant effect by targeting GPVI. An antiplatelet agent bearing antioxidative and anti-inflammatory activities without bleeding risk may have therapeutic advantage over current antiplatelet drugs for atherothrombosis.