中性粒细胞和中性粒细胞胞外诱捕网导致小鼠蛛网膜下腔出血后血管闭塞和迟发性脑缺血。
Neutrophils and Neutrophil Extracellular Traps Cause Vascular Occlusion and Delayed Cerebral Ischemia After Subarachnoid Hemorrhage in Mice.
机构信息
The Vivian L. Smith Department of Neurosurgery, McGovern Medical School (H.A.Z., S.-H.H., A.D., K.T., P.T.P., K.M., J.P.H., S.L.B., D.W.M.), University of Texas Health Science Center at Houston.
Department of Neurology, McGovern Medical School (P.P., A.C., S.P.M., L.D.M., J.A.), University of Texas Health Science Center at Houston.
出版信息
Arterioscler Thromb Vasc Biol. 2024 Mar;44(3):635-652. doi: 10.1161/ATVBAHA.123.320224. Epub 2024 Feb 1.
BACKGROUND
After subarachnoid hemorrhage (SAH), neutrophils are deleterious and contribute to poor outcomes. Neutrophils can produce neutrophil extracellular traps (NETs) after ischemic stroke. Our hypothesis was that, after SAH, neutrophils contribute to delayed cerebral ischemia (DCI) and worse outcomes via cerebrovascular occlusion by NETs.
METHODS
SAH was induced via endovascular perforation, and SAH mice were given either a neutrophil-depleting antibody, a PAD4 (peptidylarginine deiminase 4) inhibitor (to prevent NETosis), DNAse-I (to degrade NETs), or a vehicle control. Mice underwent daily neurological assessment until day 7 and then euthanized for quantification of intravascular brain NETs (iNETs). Subsets of mice were used to quantify neutrophil infiltration, NETosis potential, iNETs, cerebral perfusion, and infarction. In addition, NET markers were assessed in the blood of aneurysmal SAH patients.
RESULTS
In mice, SAH led to brain neutrophil infiltration within 24 hours, induced a pro-NETosis phenotype selectively in skull neutrophils, and caused a significant increase in iNETs by day 1, which persisted until at least day 7. Neutrophil depletion significantly reduced iNETs, improving cerebral perfusion, leading to less neurological deficits and less incidence of DCI (16% versus 51.9%). Similarly, PAD4 inhibition reduced iNETs, improved neurological outcome, and reduced incidence of DCI (5% versus 30%), whereas degrading NETs marginally improved outcomes. Patients with aneurysmal SAH who developed DCI had elevated markers of NETs compared with non-DCI patients.
CONCLUSIONS
After SAH, skull-derived neutrophils are primed for NETosis, and there are persistent brain iNETs, which correlated with delayed deficits. The findings from this study suggest that, after SAH, neutrophils and NETosis are therapeutic targets, which can prevent vascular occlusion by NETs in the brain, thereby lessening the risk of DCI. Finally, NET markers may be biomarkers, which can predict which patients with aneurysmal SAH are at risk for developing DCI.
背景
蛛网膜下腔出血（SAH）后，中性粒细胞具有危害性，并导致不良预后。缺血性卒中后中性粒细胞可产生中性粒细胞胞外诱捕网（NETs）。我们的假设是，SAH 后，通过 NET 引起的脑血管闭塞，中性粒细胞导致迟发性脑缺血（DCI）和更差的预后。
方法
通过血管内穿孔诱导 SAH，给予 SAH 小鼠中性粒细胞耗竭抗体、PAD4（肽基精氨酸脱亚氨酶 4）抑制剂（预防 NETosis）、DNAse-I（降解 NETs）或载体对照。小鼠每天进行神经学评估，直到第 7 天，然后处死以量化血管内脑 NETs（iNETs）。部分小鼠用于量化中性粒细胞浸润、NETosis 潜能、iNETs、脑灌注和梗死。此外，还评估了动脉瘤性 SAH 患者血液中的 NET 标志物。
结果
在小鼠中，SAH 在 24 小时内导致脑中性粒细胞浸润，选择性诱导颅骨中性粒细胞产生促 NETosis 表型，并在第 1 天引起明显的 iNETs 增加，至少持续到第 7 天。中性粒细胞耗竭显著减少 iNETs，改善脑灌注，导致神经功能缺损和 DCI 的发生率降低（16%对 51.9%）。同样，PAD4 抑制减少 iNETs，改善神经功能预后，降低 DCI 的发生率（5%对 30%），而降解 NETs 则适度改善了预后。与非 DCI 患者相比，发生 DCI 的动脉瘤性 SAH 患者的 NET 标志物升高。
结论
SAH 后，颅骨来源的中性粒细胞被 NETosis 所激活，且存在持续的脑内 iNETs，这与迟发性缺损相关。这项研究的结果表明，SAH 后，中性粒细胞和 NETosis 是治疗靶点，可以防止 NET 在脑中引起血管闭塞，从而降低 DCI 的风险。最后，NET 标志物可能是生物标志物，可预测哪些动脉瘤性 SAH 患者有发生 DCI 的风险。
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