Neuroprotective effect of Apelin 13 on ischemic stroke by activating AMPK/GSK-3β/Nrf2 signaling
Neuroprotective effect of Apelin 13 on ischemic stroke by activating AMPK/GSK-3β/Nrf2 signaling
复制标题
Apelin 13 通过激活 AMPK/GSK-3β/Nrf2 信号对缺血性中风的神经保护作用
DOI：
10.1186/s12974-019-1406-7
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发表时间：
2019-02-01
影响因子：
9.3
通讯作者：
Wen, Aidong
中科院分区：
文献类型：
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作者：
Duan, Jialin;Cui, Jia;Wen, Aidong
背景研究表明Apelin 13对缺血再灌注（I/R）诱导的细胞凋亡具有保护作用。然而，Apelin 13保护脑I/R的机制仍有待阐明。本研究旨在探讨Apelin 13是否通过AMPK/GSK-3/Nrf 2通路发挥脑保护作用。测定神经行为学评分、脑梗死体积及脑组织细胞因子。对于体外研究，使用PC 12细胞。为了阐明机制，使用蛋白酶抑制剂或siRNA。蛋白质水平进行了调查，通过westernblotting. ResultsApelin 13治疗的结果表明，显着减少梗死面积，改善神经功能的结果，减少脑水肿，并抑制细胞凋亡，氧化应激，和神经炎症后I/R。Apelin 13显著增加Nrf 2的表达以及AMPK和GSK-3的磷酸化水平。此外，在培养的PC 12细胞中，也观察到相同的保护作用。用其siRNA沉默Nrf 2基因消除了Apelin 13对I/R诱导的PC 12细胞损伤、氧化应激和炎症的预防。通过其siRNA抑制AMPK降低了Apelin 13诱导的Nrf 2表达水平，并减弱了Apelin 13的保护作用。GSK-3和Nrf 2之间的相互作用关系也通过相对过表达得到证实。结论Apelin 13通过AR/G/PLC/IP 3/CaMKK信号通路激活AMPK/GSK-3通路，进而上调Nrf 2调控的抗氧化酶的表达，从而保护缺血再灌注损伤后ROS介导的炎症和氧化应激。
BackgroundPrevious studies had showed that Apelin 13 could protect against apoptosis induced by ischemic/reperfusion (I/R). However, the mechanisms whereby Apelin 13 protected brain I/R remained to be elucidated. The present study was designed to determine whether Apelin 13 provided protection through AMPK/GSK-3/Nrf2 pathway.MethodsIn vivo, the I/R model was induced and Apelin 13 was given intracerebroventricularly 15min before reperfusion. The neurobehavioral scores, infarction volumes, and some cytokines in the brain were measured. For in vitro study, PC12 cells were used. To clarify the mechanisms, proteases inhibitors or siRNA were used. Protein levels were investigated by western blotting.ResultsThe results showed that Apelin 13 treatment significantly reduced infarct size, improved neurological outcomes, decreased brain edema, and inhibited cell apoptosis, oxidative stress, and neuroinflammation after I/R. Apelin 13 significantly increased the expression of Nrf2 and the phosphorylation levels of AMPK and GSK-3. Furthermore, in cultured PC12 cells, the same protective effects were also observed. Silencing Nrf2 gene with its siRNA abolished the Apelin 13's prevention of I/R-induced PC12 cell injury, oxidative stress, and inflammation. Inhibition of AMPK by its siRNA decreased the level of Apelin 13-induced Nrf2 expression and diminished the protective effects of Apelin 13. The interplay relationship between GSK-3 and Nrf2 was also verified with relative overexpression. Using selective inhibitors, we further identified the upstream of AMPK/GSK-3/Nrf2 is AR/G/PLC/IP3/CaMKK.ConclusionsIn conclusion, the previous results showed that Apelin 13 protected against I/R-induced ROS-mediated inflammation and oxidative stress through activating the AMPK/GSK-3 pathway by AR/G/PLC/IP3/CaMKK signaling, and further upregulated the expression of Nrf2-regulated antioxidant enzymes.