Increasing evidence has indicated that the Golgi apparatus (GA) is involved in the development of cerebral ischemia–reperfusion (IR) injury. Finding effective neuroprotective agents targeting GA has become a priority in the treatment of ischemic stroke. GM130, a key structural protein present on the cis-face of the GA, maintains its structure through its phosphorylation and dephosphorylation. However, the molecular mechanisms by which GM130 regulates IR-induced neuronal apoptosis are not well elucidated. Mouse neuroblastoma Neuro2a (N2A) cells were subjected to oxygen–glucose deprivation and reperfusion (OGDR) insult. Cell proliferation and apoptosis were determined using MTT assay, TUNEL staining, and flow cytometry. GA morphology was detected by immunocytochemical staining and immunofluorescence microscopy. GA-related protein and mRNA levels were detected by WB and qPCR, respectively. Treatment with Purvalanol A, an effective Cdk1 inhibitor, and transfection of Cdk1-shRNA were carried out to inhibit OGDR-induced Cdk1 elevation. The results demonstrated that OGDR induced Golgi fragmentation, neuronal apoptosis, GM130 phosphorylation, and p115 cleavage in N2A cells. Cdk1 elevation after OGDR was closely correlated with GM130 phosphorylation, not p115. Inhibition of Cdk1 significantly attenuated OGDR-induced Golgi fragmentation and cell apoptosis. Cdk1 interacted with GM130 and decreased its phosphorylation on the serine 25 site in N2A cells exposed to OGDR. The present findings reveal that Cdk1 protects against IR-induced GA fragmentation and apoptosis, likely through the mediation of GM130 phosphorylation. This neuroprotective potential of Cdk1 against IR insult and the underlying mechanism will pave the way for potential clinical applications targeting the GA organelle for cerebral IR-related disorders.

越来越多的证据表明高尔基体（GA）参与脑缺血再灌注（IR）损伤的发生。寻找针对GA的有效神经保护剂已成为治疗缺血性脑卒中的首要任务。GM130 是 GA 顺面的关键结构蛋白，通过磷酸化和去磷酸化维持其结构。然而，GM130 调节 IR 诱导的神经元凋亡的分子机制尚未得到很好的阐明。小鼠神经母细胞瘤 Neuro2a (N2A) 细胞受到氧-葡萄糖剥夺和再灌注 (OGDR) 损伤。使用MTT测定、TUNEL染色和流式细胞术测定细胞增殖和凋亡。通过免疫细胞化学染色和免疫荧光显微镜检测GA形态。分别通过WB和qPCR检测GA相关蛋白和mRNA水平。使用有效的 Cdk1 抑制剂 Purvalanol A 进行治疗并转染 Cdk1-shRNA，以抑制 OGDR 诱导的 Cdk1 升高。结果表明，OGDR 诱导 N2A 细胞中高尔基体断裂、神经元凋亡、GM130 磷酸化和 p115 裂解。OGDR 后 Cdk1 升高与 GM130 磷酸化密切相关，而不是 p115。抑制 Cdk1 显着减弱 OGDR 诱导的高尔基体断裂和细胞凋亡。在暴露于 OGDR 的 N2A 细胞中，Cdk1 与 GM130 相互作用并降低其丝氨酸 25 位点的磷酸化。目前的研究结果表明，Cdk1 可能通过 GM130 磷酸化的介导来防止 IR 诱导的 GA 断裂和细胞凋亡。Cdk1 针对 IR 损伤的神经保护潜力及其潜在机制将为针对 GA 细胞器治疗脑 IR 相关疾病的潜在临床应用铺平道路。