Mdivi-1 Protects Against Ischemic Brain Injury via Elevating Extracellular Adenosine in a cAMP/CREB-CD39-Dependent Manner
Mdivi-1 Protects Against Ischemic Brain Injury via Elevating Extracellular Adenosine in a cAMP/CREB-CD39-Dependent Manner
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Mdivi-1 通过以 cAMP/CREB-CD39 依赖性方式升高细胞外腺苷来预防缺血性脑损伤
DOI：
10.1007/s12035-014-9002-4
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发表时间：
2016-01-01
影响因子：
5.1
通讯作者：
Dong, Qiang
中科院分区：
文献类型：
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作者：
Cui, Mei;Ding, Hongyan;Dong, Qiang
本研究旨在探讨 Mdivi-1 的神经保护作用是否归因于细胞外 ATP 和腺苷。 Mdivi-1 在大脑中动脉闭塞（MCAO）之前或之后施用。在 MCAO 小鼠模型中通过体内微透析和高压液相色谱 (HPLC) 测量细胞外腺苷。进行蛋白质印迹以确定Mdivi-1对体内和培养的星形胶质细胞中CD39和CREB磷酸化表达的影响。在原代星形胶质细胞中检测到细胞内 cAMP 和蛋白激酶 A (PKA) 活性。结果显示，在 MCAO 之前或之后服用 Mdivi-1 可以显着减少梗塞体积和神经系统评分。有趣的是，Mdivi-1 预处理导致细胞外腺苷显着增加，同时 ATP 减少。 CD39（而非 CD73）的表达被 Mdivi-1 上调，Mdivi-1 与磷酸化 cAMP 反应元件结合蛋白（CREB）升高相关，CREB 是一种可能调节 CD39 表达的转录因子。在原代星形胶质细胞中，Mdivi-1 处理诱导细胞内 cAMP、PKA 活性和 CREB 磷酸化增加，而 PKA 特异性抑制剂完全逆转 Mdivi-1 诱导的 CD39 表达。我们的结果表明，Mdivi-1 通过增加细胞外腺苷来防止缺血性脑损伤，这一过程涉及 CD39 表达升高，可能受到 cAMP/PKA/CREB 级联的调节。
This study aimed to examine whether the neuroprotective effects of Mdivi-1 are attributable to extracellular ATP and adenosine. Mdivi-1 was administered prior to or post middle cerebral artery occlusion (MCAO). The extracellular adenosine was measured by in vivo microdialysis and high-pressure liquid chromatography (HPLC) in MCAO mouse model. Western blot was done to determine the influence of Mdivi-1 on the expression of CD39 and CREB phosphorylation both in vivo and in the cultured astrocytes. Intracellular cAMP and protein kinase A (PKA) activity were detected in primary astrocytes. Results showed that Mdivi-1 significantly reduced infarct volume and neurological scores when administered either prior to or post MCAO. Interestingly, pretreatment with Mdivi-1 resulted in marked increase of extracellular adenosine and concomitant decrease in ATP. The expression of CD39, but not CD73, was upregulated by Mdivi-1, which was associated with the elevated phosphorylated cAMP response element-binding protein (CREB), a transcription factor potentially regulating CD39 expression. In primary astrocytes, Mdivi-1 treatment induced increases in intracellular cAMP, PKA activity and CREB phosphorylation, and PKA-specific inhibitor completely reversed Mdivi-1-induced CD39 expression. Our results demonstrate that Mdivi-1 protects against ischemic brain injury through increasing extracellular adenosine, a process involving elevated CD39 expression that is likely modulated by cAMP/PKA/CREB cascade.