理由：中风会引起体内代谢变化，而代谢物已成为中风的潜在生物标志物。然而，中风涉及的具体代谢物以及中风期间脑损伤的机制仍不清楚。方法：对 69 名对照者和 51 名在 24 小时内接受再灌注的缺血性卒中患者的临床血清样本进行表面增强拉曼光谱 (SERS) 和液相色谱-质谱 (LC-MS) 分析，以鉴定差异丰富的代谢物。对小鼠进行短暂大脑中动脉闭塞（tMCAO），然后静脉注射次黄嘌呤。通过氯化四唑（TTC）染色评估梗塞面积，并进行行为测试。通过伊文思蓝和 IgG 染色评估血脑屏障 (BBB) 渗漏。利用人血管类器官研究次黄嘌呤诱导内皮细胞焦亡的机制。结果：SERS 和 LC-MS 以高灵敏度、快速和准确的方式揭示了中风患者和对照者血清的代谢特征。在缺血性中风的急性期，患者和小鼠的次黄嘌呤水平均显着升高（Bonferroni 校正后 p < 0.001）。此外，增加次黄嘌呤会增加缺血性中风小鼠的梗塞面积，加剧血脑屏障渗漏和神经行为缺陷。使用内皮细胞、人类血管类器官和中风小鼠进行的进一步机制研究表明，次黄嘌呤介导的gasdermin E (GSDME)依赖性内皮细胞焦亡是通过细胞内Ca2+超载发生的。结论：我们的研究确定次黄嘌呤是一种重要的代谢物，可通过触发 GSDME 依赖性内皮细胞焦亡来诱导中风中的血管损伤和 BBB 破坏。© 作者。

Rationale: Stroke induces metabolic changes in the body, and metabolites have become potential biomarkers for stroke. However, the specific metabolites involved in stroke and the mechanisms underlying brain injury during stroke remain unclear. Methods: Surface-enhanced Raman spectroscopy (SERS) and liquid chromatography-mass spectrometry (LC‒MS) analysis of clinical serum samples from 69 controls and 51 ischemic stroke patients who underwent reperfusion within 24 hours were performed to identify differentially abundant metabolites. Mice were subjected to transient middle cerebral artery occlusion (tMCAO) and then intravenously injected with hypoxanthine. The infarct area was evaluated via tetrazolium chloride (TTC) staining, and behavior tests were conducted. Blood-brain barrier (BBB) leakage was assessed by Evans blue and IgG staining. Human blood vessel organoids were used to investigate the mechanism of hypoxanthine-induced pyroptosis of endothelial cells. Results: SERS and LC‒MS revealed the metabolic profiles of serum from stroke patients and controls with high sensitivity, speed and accuracy. Hypoxanthine levels were significantly elevated in the acute stage of ischemic stroke in both patients and mice (p < 0.001 after Bonferroni correction). In addition, increasing hypoxanthine increased the infarct area and aggravated BBB leakage and neurobehavioral deficits in mice after ischemic stroke. Further mechanistic studies using endothelial cells, human blood vessel organoids, and stroke mice demonstrated that hypoxanthine-mediated gasdermin E (GSDME)-dependent pyroptosis of endothelial cells occurs through intracellular Ca2+ overload. Conclusion: Our study identified hypoxanthine as an important metabolite that induces vascular injury and BBB disruption in stroke through triggering GSDME-dependent pyroptosis of endothelial cells.© The author(s).