目的：采用生物信息学筛选治疗脑缺血的神经保护药物并在动物 tMCAO 模型中进行评价。方法：从 Gene Expression Omnibus（GEO）数据库中获得MCAO 6 h 与12 h 两组数据集后进行差异基因分析。使用 DAVID和Metascape对差异基因进行生物过程与信号通路富集分析。使用Cytoscape筛选出关键基因并在 小鼠tMCAO模型中进行验证。采用Connectivity Map（Cmap）在线工具预测潜在的脑缺血治疗药物，并选 取自噬抑制剂KU-0063794探讨对tMCAO小鼠的神经保护作用。结果：两组数据集共鉴定出289个共同差 异基因。富集分析结果表明，差异基因在缺血性脑卒中急性期主要富集于突触传递调控、脂质生物合成、细 胞凋亡等生物学过程。Real-time PCR结果显示，tMCAO后12 h 梗死周围皮质中Hmox1、Serpine1、Ptgs2、 Timp1 的 mRNA 水平均升高；Western blotting 结果表明在缺血损伤后 6～24 h，Hmox1 的表达水平增加。 KU-0063794可显著减少tMCAO小鼠脑梗死体积、并通过激活Nrf2/Hmox1通路减少氧化应激反应和神经 细胞凋亡水平；行为学测试表明，KU-0063794显著改善tMCAO小鼠的运动功能障碍。结论：KU-0063794 通过减少氧化应激反应和神经细胞凋亡发挥神经保护作用。

To employ bioinformatics to screen for neuroprotective drugs for cerebral ischemia and evaluate them in an animal transient Middle Cerebral Artery Occlusion (tMCAO) model. Methods: Two datasets from the Gene Expression Omnibus (GEO) database, representing the 6 h and 12 h time points after MCAO, were obtained and subjected to differential gene analysis. The biological processes and signaling pathways of the differentially expressed genes were enriched using DAVID and Metascape. Key genes were identified using Cytoscape and validated in a mouse tMCAO model. The Connectivity Map (Cmap) online tool was used to predict potential therapeutic drugs for cerebral ischemia, and the autophagy inhibitor KU-0063794 was selected to investigate its neuroprotective effect on tMCAO mice. Results: A total of 289 common differentially expressed genes were identified between the two datasets. Enrichment analysis revealed that the differentially expressed genes were primarily enriched in biological processes such as regulation of synaptic transmission, lipid biosynthesis, and apoptosis during the acute phase of ischemic stroke. Real-time PCR results showed that the mRNA levels of Hmox1, Serpine1, Ptgs2, and Timp1 in the peri-infarct cortex increased 12 h after tMCAO. Western blotting results indicated that the expression level of Hmox1 increased from 6 to 24 h after ischemic injury. KU-0063794 significantly reduced cerebral infarct volume in tMCAO mice, mitigated oxidative stress and neuronal apoptosis by activating the Nrf2/Hmox1 pathway, and behavioral tests demonstrated that KU-0063794 significantly improved motor dysfunction in tMCAO mice. Conclusion: KU-0063794 exerts neuroprotective effects by reducing oxidative stress and neuronal apoptosis.