MicroRNA-139-5p negatively regulates NR2A-containing NMDA receptor in the rat pilocarpine model and patients with temporal lobe epilepsy
MicroRNA-139-5p negatively regulates NR2A-containing NMDA receptor in the rat pilocarpine model and patients with temporal lobe epilepsy
复制标题
MicroRNA-139-5p在大鼠毛果芸香碱模型和颞叶癫痫患者中负向调节含NR2A的NMDA受体
DOI：
10.1111/epi.13568
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发表时间：
2016-11-01
期刊：
影响因子：
5.6
通讯作者：
Li, Jing
中科院分区：
文献类型：
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作者：
Alsharafi, Walid A.;Xiao, Bo;Li, Jing
目的癫痫持续状态（SE）时N-甲基-d-天冬氨酸（NMDA）亚基NR 2A和NR 2B表达的调控机制尚不完全清楚。在这里，我们探讨了脑富集microRNA（miR）-139-5p在这一过程中的作用。MethodsmiRNA微阵列进行检查的变化，在大鼠毛果芸香碱模型NMDA受体阻断后的miRNA表达。采用定量逆转录聚合酶链反应（qRT-PCR）和蛋白质印迹法（Western blot）检测大鼠颞叶癫痫（TLE）发展3个阶段中miR-139- 5 p、NR 2A和NR 2B水平的动态表达模式。类似的表达方法应用于从TLE患者和正常对照中获得的campi。此外，miR-139- 5 p阿戈米尔和阿托米尔被用来探讨miR-139- 5 p在确定NMDA受体亚基的表达patterns.ResultsWe确定了18个miRNA的作用，显着改变了大鼠毛果芸香碱模型后NMDA受体阻滞。其中，miR-139- 5 p显著上调，并且预测Grin 2A是其潜在的推定靶点。在TLE患者中，miR-139- 5 p表达显著下调，而NR 2A和NR 2B水平显著上调。在SE大鼠模型中，miR-139- 5 p在急性期和慢性期表达下调，NR 2A表达上调，但在潜伏期无明显变化。NR 2B的表达在TLE发育的三个阶段均上调。miR-139- 5 p的过表达降低，而miR-139- 5 p的缺失增强了由毛果芸香碱处理诱导的NR 2A的表达水平，而不是NR 2B。有趣的是，NMDA非选择性拮抗剂和NR 2A选择性拮抗剂增强了毛果芸香碱治疗抑制的miR-139- 5 p水平，而NR 2B选择性拮抗剂则无效。重要性这些发现阐明了miR-139- 5 p在TLE发展中的NMDA受体参与中的潜在作用，并可能为TLE的未来治疗提供新的治疗靶点。
ObjectivesRegulation of N-methyl-d-aspartate (NMDA) subunits NR2A and NR2B expression during status epilepticus (SE) remains incompletely understood. Here we explored the role of brain-enriched microRNA (miR)-139-5p in this process.MethodsmiRNA microarray was performed to examine changes in miRNA expression in the rat pilocarpine model following NMDA-receptor blockade. The dynamic expression patterns of miR-139-5p, NR2A, and NR2B levels were measured in rats during the three phases of temporal lobe epilepsy (TLE) development using quantitative reverse transcription polymerase chain reaction (qRT-PCR) and Western blot. Similar expression methods were applied to hippocampi obtained from patients with TLE and from normal controls. Moreover, miR-139-5p agomir and antagomir were utilized to explore the role of miR-139-5p in determining NMDA-receptor subunit expression patterns.ResultsWe identified 18 miRNAs that were significantly altered in the rat pilocarpine model following NMDA-receptor blockade. Of these, miR-139-5p was significantly up-regulated and Grin2A was predicted as its potential putative target. In patients with TLE, miR-139-5p expression was significantly down-regulated, whereas NR2A and NR2B levels were significantly up-regulated. In the rat model of SE, miR-139-5p expression was down-regulated while NR2A was up-regulated in the acute and chronic phases, but not in the latent phase. NR2B expression was up-regulated during the three phases of TLE development. Overexpression of miR-139-5p decreased, whereas depletion of miR-139-5p enhanced the expression levels of NR2A, but not NR2B, induced by pilocarpine treatment. Of interest, NMDA nonselective antagonist and NR2A selective antagonist enhanced miR-139-5p levels suppressed by pilocarpine treatment, whereas the NR2B selective antagonist was ineffective.SignificanceThese findings elucidate the potential role of miR-139-5p in NMDA-receptor involvement in TLE development and may provide novel therapeutic targets for the future treatment of TLE.