Jiahang Sun1, Xiaoying Gao2, Dawei Meng3, Yang Xu4, Xichun Wang5, Xin Gu6, Mian Guo1, Xiaodong Shao7, Hongwen Yan8, Chuanlu Jiang1, Yongri Zheng1. 1. Department of Neurosurgery, The Second Affiliated Hospital of Harbin Medical University, Harbin, China. 2. Department of Anesthesiology, The Fourth Affiliated Hospital of Harbin Medical University, Harbin, China. 3. Department of Neurosurgery, Aviation General Hospital of China Medical University, Beijing, China. 4. Department of Urology, Cancer Hospital of Harbin Medical University, Harbin, China. 5. Department of Neurosurgery, Heilongjiang Provincial Hospital, Harbin, China. 6. Department of Head and Neck Surgery, Cancer Hospital of Harbin Medical University, Harbin, China. 7. School of Basic Medical Sciences, Harbin Medical University, Harbin, China. 8. Department of Pediatric Hematology, Peking University International Hospital, Beijing, China.
Abstract
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.
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.
Authors: Amaya Sanz Rodriguez; Tobias Engel; Arpad Palfi; G Jane Farrar; David C Henshall; Eva M Jimenez-Mateos Journal: Int J Physiol Pathophysiol Pharmacol Date: 2017-12-25
Authors: Gareth Morris; Gary P Brennan; Cristina R Reschke; David C Henshall; Stephanie Schorge Journal: Epilepsia Date: 2018-07-05 Impact factor: 5.864