K6PC-5 Activates SphK1-Nrf2 Signaling to Protect Neuronal Cells from Oxygen Glucose Deprivation/Re-Oxygenation
K6PC-5 Activates SphK1-Nrf2 Signaling to Protect Neuronal Cells from Oxygen Glucose Deprivation/Re-Oxygenation
复制标题
K6PC-5 激活 SphK1-Nrf2 信号传导以保护神经元细胞免受缺氧/再氧合的影响
DOI：
10.1159/000495716
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发表时间：
2018-01-01
影响因子：
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通讯作者：
Di, Guangfu
中科院分区：
文献类型：
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作者：
Liu, Hua;Zhang, Zhiqing;Di, Guangfu
背景/目的：需要新的策略来对抗神经元缺血-再灌注损伤。K6PC-5是一种新的鞘氨醇激活剂，其在神经细胞中的潜在活性尚未被检测。方法：分别用细胞计数试剂盒-8法和乳酸脱氢酶释放法检测细胞存活和坏死情况。用JC-1染色法检测线粒体去极化。用实时定量聚合酶链式反应和Western blotting检测核因子红系相关因子2(Nrf2)信号成分的表达水平。结果：K6PC-5对SH-SY5Y神经元和原代培养的小鼠海马神经元具有保护作用。K6PC-5激活SphK1，靶向短发夹状RNA(ShRNA)敲除SphK1几乎完全取消K6PC-5诱导的神经细胞保护作用。进一步的研究表明，K6PC-5能抑制OGDR诱导的神经细胞程序性坏死。重要的是，K6PC-5激活了SphK1下游的Nrf2信号。靶向shRNA沉默Nrf2几乎完全无效K6PC-5介导的神经细胞保护作用。结论：K6PC-5激活SphK1-Nrf2信号通路，保护神经细胞免受OGDR的损伤。K6PC-5可能是一种有前途的脑缺血再灌注损伤的神经保护策略。
Background/Aims: New strategies are required to combat neuronal ischemia-reperfusion injuries. K6PC-5 is a novel sphingosine kinase 1 (SphK1) activator whose potential activity in neuronal cells has not yet been tested. Methods: Cell survival and necrosis were assessed with a Cell Counting Kit-8 assay and lactate dehydrogenase release assay, respectively. Mitochondrial depolarization was tested by a JC-1 dye assay. Expression levels of nuclear factor erythroid 2-related factor 2 (Nrf2) signaling components were examined by quantitative real-timePCR and western blotting. Results: K6PC-5 protected SH-SY5Y neuronal cells and primary murine hippocampal neurons from oxygen glucose deprivation/re-oxygenation (OGDR). K6PC-5 activated SphK1, and SphK1 knockdown by targeted short hairpin RNA (shRNA) almost completely abolished K6PC-5-induced neuronal cell protection. Further work showed that K6PC-5 inhibited OGDR-induced programmed necrosis in neuronal cells. Importantly, K6PC-5 activated Nrf2 signaling, which is downstream of SphK1. Silencing of Nrf2 by targeted shRNA almost completely nullified K6PC-5-mediated neuronal cell protection against OGDR. Conclusion: K6PC-5 activates SphK1-Nrf2 signaling to protect neuronal cells from OGDR. K6PC-5 might be a promising neuroprotective strategy for ischemia-reperfusion injuries.