A novel GSK-3β inhibitor YQ138 prevents neuronal injury induced by glutamate and brain ischemia through activation of the Nrf2 signaling pathway
A novel GSK-3β inhibitor YQ138 prevents neuronal injury induced by glutamate and brain ischemia through activation of the Nrf2 signaling pathway
复制标题
新型 GSK-3β 抑制剂 YQ138 通过激活 Nrf2 信号通路预防谷氨酸和脑缺血引起的神经元损伤
DOI：
10.1038/aps.2016.3
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发表时间：
2016-06-01
影响因子：
8.2
通讯作者：
Li, Jia
中科院分区：
文献类型：
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作者：
Pang, Tao;Wang, Yun-jie;Li, Jia
目的：发现神经保护化合物，并表征所发现的活性化合物YQ138作为一种新型GSK-3β抑制剂。方法：用谷氨酸处理原代大鼠小脑颗粒细胞（CGCs），并用MTT法分析细胞活力，作为筛选神经保护化合物的体外模型。在 OGD 或血清剥夺诱导的神经元损伤模型中进一步测试了活性化合物。采用Western blotting检测GSK-3β下游蛋白（Nrf2、HO-1、NQO1、Tau、β-catenin）的表达水平。为了评估体内神经保护作用，对成年雄性大鼠进行短暂大脑中动脉闭塞（tMCAO），然后在缺血发作后2、4和6小时用YQ138（10 mg/kg，静脉注射）治疗。结果：从由约2000种潜在激酶抑制剂组成的化合物库中，发现YQ138发挥神经保护作用 作用：YQ138（0.1-40 μmol/L）预处理可剂量依赖性地抑制谷氨酸诱导的神经元死亡。此外，YQ138 (10 μmol/L) 预处理显着抑制 OGD 或血清剥夺诱导的神经元死亡。在测试的七种激酶中，YQ138 选择性抑制 GSK-3β 的活性（IC 50 = 0.52 nmol/L）。此外，YQ138剂量依赖性地增加CGC中β-连环蛋白的表达，并降低Tau的磷酸化。此外，YQ138显着增加CGC中GSK-3β下游抗氧化蛋白Nrf2、HO-1、NQO1、GSH和SOD的表达。在tMCAO大鼠中，给予YQ138可显着减少梗塞体积，改善神经功能缺损，并增加大脑皮层Nrf2和HO-1的表达以及SOD和GSH的活性。结论：新型GSK-3β抑制剂YQ138在体外和体内均能有效抑制脑缺血损伤。
Aim:To discover neuroprotective compounds and to characterize the discovered active compound YQ138 as a novel GSK-3β inhibitor.Methods:Primary rat cerebellar granule cells (CGCs) were treated with glutamate, and cell viability was analyzed with MTT assay, which was used as in vitro model for screening neuroprotective compounds. Active compound was further tested in OGD-or serum deprivation-induced neuronal injury models. The expression levels of GSK-3β downstream proteins (Nrf2, HO-1, NQO1, Tau and β-catenin) were detected with Western blotting. For evaluating the neuroprotective effects in vivo, adult male rats were subjected to transient middle cerebral artery occlusion (tMCAO), then treated with YQ138 (10 mg/kg, iv) at 2, 4 and 6 h after ischemia onset.Results:From a compound library consisting of about 2000 potential kinase inhibitors, YQ138 was found to exert neuroprotective effects: pretreatment with YQ138 (0.1–40 μmol/L) dose-dependently inhibited glutamate-induced neuronal death. Furthermore, pretreatment with YQ138 (10 μmol/L) significantly inhibited OGD-or serum deprivation-induced neuronal death. Among a panel of seven kinases tested, YQ138 selectively inhibited the activity of GSK-3β (IC 50= 0.52 nmol/L). Furthermore, YQ138 dose-dependently increased the expression of β-catenin, and decreased the phosphorylation of Tau in CGCs. Moreover, YQ138 significantly increased the expression of GSK-3β downstream antioxidative proteins Nrf2, HO-1, NQO1, GSH and SOD in CGCs. In rats with tMCAO, administration of YQ138 significantly decreased infarct volume, improved the neurological deficit, and increased the expression of Nrf2 and HO-1 and the activities of SOD and GSH in the cerebral cortex.Conclusion:A novel GSK-3β inhibitor YQ138 effectively suppresses brain ischemic injury in vitro and in vivo.