Department of Critical Medicine, The Second Affiliated Hospital of Shandong First Medical University, Taian 271000, China.
Department of Emergency, The Second Affiliated Hospital of Shandong First Medical University, Taian 271000, China.
Neuroscience. 2023 Aug 21;526:175-184. doi: 10.1016/j.neuroscience.2023.06.024. Epub 2023 Jul 3.
Brain injury represents a leading cause of deaths following cardiac arrest (CA) and cardiopulmonary resuscitation (CPR). This study explores the role of CREB1 (cAMP responsive element binding protein 1)/DAPK1 (death associated protein kinase 1) axis in brain injury after CPR. CA was induced by asphyxia in rats, followed by CPR. After CREB1 over-expression, the survival rate and neurological function score of rats were measured. Nissl and TUNEL staining evaluated the pathological condition of hippocampus and apoptosis of hippocampal neurons respectively. H19-7 cells were subjected to OGD/R and infected with oe-CREB1. CCK-8 assay and flow cytometry measured the cell viability and apoptosis. CREB1, DAPK1, and cleaved Caspase-3 expressions were examined using Western blot. The binding between CREB1 and DAPK1 was determined using ChIP and dual-luciferase reporter assays. CREB1 was poorly expressed while DAPK1 was highly expressed in rat hippocampus after CPR. CREB1 overexpression improved rat neurological function, repressed neuron apoptosis, and reduced cleaved Caspase-3 expression. CREB1 was enriched on the DAPK1 promoter and suppressed DAPK1 expression. DAPK1 overexpression reversed the inhibition of OGD/R-insulted apoptosis by CREB1 overexpression. To conclude, CREB1 suppresses hippocampal neuron apoptosis and mitigates brain injury after CPR by inhibiting DAPK1 expression.
脑损伤是心脏骤停(CA)和心肺复苏(CPR)后死亡的主要原因。本研究探讨了 CREB1(cAMP 反应元件结合蛋白 1)/DAPK1(凋亡相关蛋白激酶 1)轴在 CPR 后脑损伤中的作用。通过窒息在大鼠中诱导 CA,然后进行 CPR。在过表达 CREB1 后,测量大鼠的存活率和神经功能评分。Nissl 和 TUNEL 染色分别评估海马的病理状况和海马神经元的凋亡。将 H19-7 细胞进行 OGD/R 处理并感染 oe-CREB1。CCK-8 测定法和流式细胞术分别测量细胞活力和细胞凋亡。使用 Western blot 检测 CREB1、DAPK1 和 cleaved Caspase-3 的表达。使用 ChIP 和双荧光素酶报告基因测定来确定 CREB1 和 DAPK1 之间的结合。CPR 后大鼠海马中 CREB1 表达水平降低,DAPK1 表达水平升高。过表达 CREB1 可改善大鼠的神经功能,抑制神经元凋亡,降低 cleaved Caspase-3 的表达。CREB1 被富集在 DAPK1 启动子上,并抑制 DAPK1 的表达。DAPK1 过表达逆转了 CREB1 过表达对 OGD/R 损伤诱导的凋亡的抑制作用。综上所述,CREB1 通过抑制 DAPK1 的表达,抑制海马神经元凋亡并减轻 CPR 后脑损伤。