Department of Neurosurgery, the Second Affiliated Hospital of Dalian Medical University, Dalian, China.
Epileptic Center of Liaoning, the Second Affiliated Hospital of Dalian Medical University, Dalian, China.
Brain Behav. 2021 Jul;11(7):e02195. doi: 10.1002/brb3.2195. Epub 2021 May 24.
Accumulating evidence demonstrates that certain microRNAs play critical roles in epileptogenesis. Our previous studies found microRNA (miR)-129-2-3p was induced in patients with refractory temporal lobe epilepsy (TLE). In this study, we aimed to explore the role of miR-129-2-3p in TLE pathogenesis.
By bioinformatics, we predicted miR-129-2-3p may target the gene GABRA1 encoding the GABA type A receptor subunit alpha 1. Luciferase assay was used to investigate the regulation of miR-129-2-3p on GABRA1 3'UTR. The dynamic expression of miR-129-2-3p and GABRA1 mRNA and protein levels were measured in primary hippocampal neurons and a rat kainic acid (KA)-induced seizure model by quantitative reverse transcription-polymerase chain reaction (qPCR), Western blotting, and immunostaining. MiR-129-2-3p agomir and antagomir were utilized to explore their role in determining GABRA1 expression. The effects of targeting miR-129-2-3p and GABRA1 on epilepsy were assessed by electroencephalography (EEG) and immunostaining.
Luciferase assay, qPCR, and Western blot results suggested GABRA1 as a direct target of miR-129-2-3p. MiR-129-2-3p level was significantly upregulated, whereas GABRA1 expression downregulated in KA-treated rat primary hippocampal neurons and KA-induced seizure model. In vivo knockdown of miR-129-2-3p by antagomir alleviated the seizure-like EEG findings in accordance with the upregulation of GABRA1. Furthermore, the seizure-suppressing effect of the antagomir was partly GABRA1 dependent.
The results suggested GABRA1 as a target of miR-129-2-3p in rat primary hippocampal neurons and a rat kainic acid (KA) seizure model. Silencing of miR-129-2-3p exerted a seizure-suppressing effect in rats. MiR-129-2-3p/GABRA1 pathway may represent a potential target for the prevention and treatment of refractory epilepsy.
越来越多的证据表明，某些 microRNAs 在癫痫发生中发挥着关键作用。我们之前的研究发现，miR-129-2-3p 在难治性颞叶癫痫（TLE）患者中被诱导。在这项研究中，我们旨在探讨 miR-129-2-3p 在 TLE 发病机制中的作用。
通过生物信息学，我们预测 miR-129-2-3p 可能靶向编码 GABA 型 A 受体亚单位α1 的基因 GABRA1。通过荧光素酶测定来研究 miR-129-2-3p 对 GABRA1 3'UTR 的调控。通过定量逆转录聚合酶链反应（qPCR）、Western 印迹和免疫染色来测量原代海马神经元和大鼠红藻氨酸（KA）诱导的癫痫模型中 miR-129-2-3p 和 GABRA1 mRNA 和蛋白水平的动态表达。使用 miR-129-2-3p agomir 和 antagomir 来探讨它们在决定 GABRA1 表达中的作用。通过脑电图（EEG）和免疫染色来评估靶向 miR-129-2-3p 和 GABRA1 对癫痫的影响。
荧光素酶测定、qPCR 和 Western blot 结果表明 GABRA1 是 miR-129-2-3p 的直接靶标。在 KA 处理的大鼠原代海马神经元和 KA 诱导的癫痫模型中，miR-129-2-3p 水平显著上调，而 GABRA1 表达下调。体内通过 antagomir 下调 miR-129-2-3p 可减轻类似癫痫的 EEG 发现，同时 GABRA1 上调。此外，antagomir 的抗癫痫作用部分依赖于 GABRA1。
结果表明 GABRA1 是大鼠原代海马神经元和大鼠红藻氨酸（KA）癫痫模型中 miR-129-2-3p 的靶标。沉默 miR-129-2-3p 在大鼠中表现出抗癫痫作用。miR-129-2-3p/GABRA1 通路可能成为预防和治疗难治性癫痫的潜在靶点。
