Department of Neurosurgery, Chongqing Medical University, Yongchuan Hospital, Yongchuan, Chongqing, China.
Department of Neurosurgery, School of Medicine, The Second Affiliated Hospital, Zhejiang University, Hangzhou, Zhejiang, China.
Neurotherapeutics. 2021 Jul;18(3):1905-1921. doi: 10.1007/s13311-021-01066-x. Epub 2021 Jun 4.
Subarachnoid hemorrhage (SAH) is a devastating cerebrovascular disease. Neuronal apoptosis plays an important pathological role in early brain injury after SAH. Galanin receptor 1 (GalR1) activation was recently shown to be anti-apoptotic in the setting of ischemic stroke. This study aimed to explore the anti-neuronal apoptosis effect of GalR1 activation after SAH, as well as the underlying mechanisms. GalR1 CRISPR and GalR1 selective agonist, M617, was administered, respectively. Extracellular-signal-regulated kinase (ERK) inhibitor (U0126) and glycogen synthase kinase 3-beta (GSK3-β) CRISPR were administered to investigate the involvement of the ERK/GSK3-β pathway in GalR1-mediated neuroprotection after SAH. Outcome assessments included neurobehavioral tests, western blot, and immunohistochemistry. The results showed that endogenous ligand galanin (Gal) and GalR1 were markedly increased in the ipsilateral brain hemisphere at 12 h and 24 h after SAH. GalR1 were expressed mainly in neurons, but expression was also observed in some astrocytes and microglia. GalR1 CRISPR knockdown exacerbated neurological deficits and neuronal apoptosis 24 h after SAH. Moreover, activation of GalR1 with M617 significantly improved short- and long-term neurological deficits but decreased neuronal apoptosis after SAH. Furthermore, GalR1 activation dysregulated the protein levels of phosphorylated ERK and GSK-3β, but downregulated the phosphorylated Tat-interactive protein 60 (TIP60) and cleaved caspase-3 at 24 h after SAH. GalR1 CRISPR, U0126, and GSK-3β CRISPR abolished the beneficial effects of GalR1 activation at 24 h after SAH in rats. Collectively, the present study demonstrated that activation of GalR1 using M617 attenuated neuronal apoptosis through the ERK/GSK-3β/TIP60 pathway after SAH in rats. GalR1 may serve as a promising therapeutic target for SAH patients.
蛛网膜下腔出血 (SAH) 是一种破坏性的脑血管疾病。神经元凋亡在 SAH 后早期脑损伤中发挥重要的病理作用。最近的研究表明，甘丙肽受体 1 (GalR1) 的激活在缺血性中风中具有抗凋亡作用。本研究旨在探讨 SAH 后 GalR1 激活的抗神经元凋亡作用及其潜在机制。分别给予 GalR1 CRISPR 和 GalR1 选择性激动剂 M617。给予细胞外信号调节激酶 (ERK) 抑制剂 (U0126) 和糖原合成酶激酶 3-β (GSK3-β) CRISPR，以研究 ERK/GSK3-β 通路在 SAH 后 GalR1 介导的神经保护中的作用。结果评估包括神经行为学测试、western blot 和免疫组织化学。结果显示，SAH 后 12 h 和 24 h，同侧大脑半球中内源性配体甘丙肽 (Gal) 和 GalR1 明显增加。GalR1 主要表达于神经元，但也在一些星形胶质细胞和小胶质细胞中表达。GalR1 CRISPR 敲低加重了 SAH 后 24 h 的神经功能缺损和神经元凋亡。此外，M617 激活 GalR1 可显著改善 SAH 后的短期和长期神经功能缺损，但减少神经元凋亡。此外，GalR1 激活可调节 ERK 和 GSK-3β 磷酸化蛋白水平，但下调 TIP60 和 cleaved caspase-3 的磷酸化 Tat 相互作用蛋白 60 (TIP60) 和 cleaved caspase-3 在 SAH 后 24 h。GalR1 CRISPR、U0126 和 GSK-3β CRISPR 消除了 GalR1 激活在 SAH 后 24 h 对大鼠的有益作用。总之，本研究表明，M617 激活 GalR1 通过 ERK/GSK-3β/TIP60 通路减轻了大鼠 SAH 后的神经元凋亡。GalR1 可能成为 SAH 患者有希望的治疗靶点。
