目的  探讨螯合锌对小鼠脑缺血急性期神经元焦亡的改善作用。方法  利用30只成年雄性昆明小鼠（25~30 g），按照随机对照的原则分为假手术组（sham）、大脑中动脉闭塞（MCAO）再灌注3 d组（MCAO）及螯合锌组（TPEN），每组10只。各组小鼠行吸入式麻醉，采用线栓法制备MCAO模型，每组小鼠选取缺血1 h再灌注3 d，螯合锌组于再灌注即刻给予腹腔注射锌离子螯合剂TPEN（10 mg/kg），sham组仅进行血管钝性分离。神经功能学评分、2,3,5-三苯基氯化四氮唑（TTC）染色、激光散斑成像评估螯合锌对小鼠脑损伤后的保护作用。透射电子显微术、免疫荧光染色、锌探针、Western blotting观察螯合锌对小鼠脑损伤后神经元焦亡的改善情况。转录组测序基因本体论（GO）富集分析，免疫荧光共染色、Western blotting观察螯合锌对黑色素瘤缺乏因子2 (AIM2)、凋亡相关斑点样蛋白 (ASC)、Caspase-1、裂解（cleaved）-Caspase-1、gasdermin(GSDM)家族蛋白GSDMD-N蛋白表达水平。结果  螯合锌改善小鼠神经功能学评分 (P<0.05)，减少小鼠脑梗死体积(P<0.01)。恢复小鼠脑血流灌注量 (P<0.001)。免疫荧光共色结果发现，与模型组比较，螯合锌能够减少神经元内锌离子聚集，减少脑缺血半球的神经元死亡数量，减少游离的锌离子含量。透射电子显微术结果发现，与模型组相比，螯合锌组小鼠脑皮质区的焦亡小体明显减少；通过转录组测序GO富集分析发现脑缺血后的炎症反应以及焦亡大规模爆发。结合Western blotting和免疫荧光结果显示，给予TPEN处理可降低脑缺血损伤引起的AIM2炎症小体蛋白表达 (P<0.05)。结论  在脑缺血急性期，螯合锌对小鼠脑缺血后细胞焦亡介导的损伤具有保护作用。

Objective  To explore the ameliorative effect of chelated zinc on neuronal pyroptosis during the acute phase of cerebral ischemia in mice. Methods  Thirty adult male Kunming mice (25-30 g) were randomly divided into three groups according to the randomized controlled principle, sham operation group (sham), middle cerebral artery occlusion (MCAO) reperfusion 3 days group (MCAO), and zinc chelation Tetrakis-(2-pyridylmethyl) ethylenediamine (TPEN) group, with 10 mice in each group. Mice in each group were anesthetized by inhalation, and MCAO model was established by the suture-occlusion method. Mice in each group were subjected to 1 hour of ischemia followed by 3 days of reperfusion. Mice in the TPEN group were intraperitoneally injected with the zinc ion chelator TPEN at a dose of 10 mg/kg immediately after reperfusion, while mice in the sham group only underwent blunt separation of blood vessels. Neurological function scoring, 2,3,5-triphenyltetrazolium chloride (TTC) staining, and laser speckle imaging were used to evaluate the protective effect of zinc chelation on cerebral injury in mice. Transmission electron microscopy, immunofluorescent staining, zinc probe, and Western blotting were performed to observe the improvement effect of zinc chelation on neuronal pyroptosis after cerebral injury in mice. Transcriptome sequencing combined with Gene Ontology (GO) enrichment analysis, immunofluorescence co-staining, and Western blotting were used to detect the expression levels of absent in melanoma 2 (AIM2), apoptosis-associated speck-like protein containing a CARD (ASC), Caspase-1, cleaved-Caspase-1, and gasdermin (GSDM) family proteins GSDMD-N proteins after zinc chelation intervention. Results  Zinc chelation improved the neurological function scores of mice (P<0.05), reduced the cerebral infarct volume (P<0.01), and restored the cerebral blood perfusion (P<0.001). Immunofluorescence co-staining results showed that compared with the model group, zinc chelation reduced intracellular zinc ion accumulation in neurons, decreased the number of neuronal deaths in the ischemic hemisphere, and lowered the content of free zinc ions. Transmission electron microscopy results revealed a significant reduction in pyroptotic bodies in the cerebral cortex of mice in the zinc chelation group compared with the model group. Transcriptome sequencing with GO enrichment analysis indicated that inflammation and massive pyroptosis erupted on 3 days after cerebral ischemia. Western blotting and immunofluorescence results demonstrated that TPEN treatment reduced the expression of AIM2 inflammasome proteins induced by cerebral ischemic injury (P<0.05). Conclusion  Chelated zinc exerts a neuroprotection against neuronal pyroptosis in mice after acute cerebral ischemia.