Time-of-Day Dependent Neuronal Injury After Ischemic Stroke: Implication of Circadian Clock Transcriptional Factor Bmal1 and Survival Kinase AKT
Time-of-Day Dependent Neuronal Injury After Ischemic Stroke: Implication of Circadian Clock Transcriptional Factor Bmal1 and Survival Kinase AKT
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DOI：
10.1007/s12035-017-0524-4
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发表时间：
2018-03-01
影响因子：
5.1
通讯作者：
Kilic, Ertugrul
中科院分区：
文献类型：
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作者：
Beker, Mustafa Caglar;Caglayan, Berrak;Kilic, Ertugrul
中风病例的发生在人类中显示出一天中的时间变化。然而，缺血性卒中后，昼夜节律与病理生理事件的内在反应机制之间的联系在很大程度上仍不清楚。为此，在小鼠中研究了对缺血/再灌注（I/R）损伤的敏感性的时间变化，其中在4个不同的Zeitgeber时间点以6小时间隔（ZT 0、ZT 6、ZT 12和ZT 18）诱导缺血性中风。除了梗死体积和脑肿胀，神经元存活，细胞凋亡，缺血，昼夜节律相关的蛋白质进行了研究，使用免疫组化，蛋白质印迹，平面表面免疫分析，液相色谱-质谱工具。在这里，我们提出的证据表明，午夜（ZT 18; 24：00）I/R损伤小鼠导致显著改善梗死体积，脑肿胀，神经功能缺损评分，神经元存活，并减少凋亡细胞死亡相比，在其他时间点诱导的缺血，这与昼夜节律蛋白Bmal 1，PerI，时钟蛋白和生存激酶AKT和Erk-1/2的表达增加。此外，核糖体蛋白S6、mTOR和Bad也显著增加，而与ZT 0（06：00）相比，在该时间点，AKT和mTOR的负调节因子PRAS 40和磷酸化p53的水平降低。此外，详细的蛋白质组学分析显示，与清晨相比，午夜时CSKP、HBB-1/2和HBA水平显著降低，而GNAZ、NEGR 1、IMPCT和PDE 1B水平升高。我们的研究结果表明，夜间I/R损伤导致不太严重的神经元损伤，增加神经元存活，增加生存激酶和昼夜节律钟蛋白的水平，并改变昼夜节律相关蛋白。
Occurrence of stroke cases displays a time-of-day variation in human. However, the mechanism linking circadian rhythm to the internal response mechanisms against pathophysiological events after ischemic stroke remained largely unknown. To this end, temporal changes in the susceptibility to ischemia/reperfusion (I/R) injury were investigated in mice in which the ischemic stroke induced at four different Zeitgeber time points with 6-h intervals (ZT0, ZT6, ZT12, and ZT18). Besides infarct volume and brain swelling, neuronal survival, apoptosis, ischemia, and circadian rhythm related proteins were examined using immunohistochemistry, Western blot, planar surface immune assay, and liquid chromatography-mass spectrometry tools. Here, we present evidence that midnight (ZT18; 24:00) I/R injury in mice resulted in significantly improved infarct volume, brain swelling, neurological deficit score, neuronal survival, and decreased apoptotic cell death compared with ischemia induced at other time points, which were associated with increased expressions of circadian proteins Bmal1, PerI, and Clock proteins and survival kinases AKT and Erk-1/2. Moreover, ribosomal protein S6, mTOR, and Bad were also significantly increased, while the levels of PRAS40, negative regulator of AKT and mTOR, and phosphorylated p53 were decreased at this time point compared to ZT0 (06:00). Furthermore, detailed proteomic analysis revealed significantly decreased CSKP, HBB-1/2, and HBA levels, while increased GNAZ, NEGR1, IMPCT, and PDE1B at midnight as compared with early morning. Our results indicate that nighttime I/R injury results in less severe neuronal damage, with increased neuronal survival, increased levels of survival kinases and circadian clock proteins, and also alters the circadian-related proteins.