Antagomirs Targeting MicroRNA-134 Increase Limk1 Levels After Experimental Seizures in Vitro and in Vivo
Antagomirs Targeting MicroRNA-134 Increase Limk1 Levels After Experimental Seizures in Vitro and in Vivo
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靶向 MicroRNA-134 的 Antagomirs 在体外和体内实验性癫痫发作后增加 Limk1 水平
DOI：
10.1159/000480647
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发表时间：
2017-01-01
影响因子：
--
通讯作者：
Zheng, Yongri
中科院分区：
文献类型：
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作者：
Sun, Jiahang;Gao, Xiaoying;Zheng, Yongri
背景：MIR-134富含于海马神经元的树突中，在癫痫的发生发展中起着重要作用。本研究旨在探讨针对miroRNA-134(Ant-134)的抗组体在实验性癫痫发作中对LIMK1表达及与miR-134结合的影响。方法：采用氯化锂-匹罗卡品建立癫痫持续状态(SE)大鼠模型，侧脑室注射Ant-134治疗。采用低镁暴露的原代神经元作为SE的体外模型。实时荧光定量聚合酶链式反应检测miR-134的表达。Western blotting检测LIMK1和Cofilin的蛋白表达。荧光素酶报告实验检测miR-134与LIMK13‘-非翻译区的结合。结果：低镁暴露组大鼠海马神经元miR-134表达明显增强。Ant-134可增加SE海马区和低镁暴露神经元LIMK1的表达，降低Cofilin的表达。此外，荧光素酶报告实验证实miR-134与LIMK1 3‘-UTR结合。MIR-134过表达抑制神经元LIMK1基因和蛋白的表达。结论：阻断miR-134可上调SE大鼠海马区LIMK1的表达，下调Cofilin的表达。这一机制可能与Ant-134的神经保护作用有关。
Background: MiR-134 is enriched in dendrites of hippocampal neurons and plays crucial roles in the progress of epilepsy. The present study aims to investigate the effects of antagomirs targeting miroRNA-134 (Ant-134) on limk1 expression and the binding of miR-134 and limk1 in experimental seizure. Methods: Status epilepticus (SE) rat model was established by lithium chloride-pilocarpine injection and was treated with Ant-134 by intracerebroventricular injection. Low Mg2+-exposed primary neurons were used as an in vitro model of SE. The expression of miR-134 was determined using real-time PCR. Protein expressions of limk1 and cofilin were determined by Western blotting. Luciferase reporter assay was used to examine the binding between miR-134 and limk1 3’-untranslated region. Results: The expression of miR-134 was markedly enhanced in hippocampus of the SE rats and low Mg2+-exposed neurons. Ant-134 increased the expression of limk1 and reduced the expression of cofilin in the SE hippocampus and Low Mg2+-exposed neurons. In addition, luciferase reporter assay confirmed that miR-134 bound limk1 3’-UTR. MiR-134 overexpression inhibited limk1 mRNA and protein expressions in neurons. Conclusion: Blockage of miR-134 upregulates limk1 expression and downregulated cofilin expression in hippocampus of the SE rats. This mechanism may contribute to the neuroprotective effects of Ant-134.