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11 November 2019 : Laboratory Research  

MicroRNA-27a-3p Downregulation Inhibits Inflammatory Response and Hippocampal Neuronal Cell Apoptosis by Upregulating Mitogen-Activated Protein Kinase 4 (MAP2K4) Expression in Epilepsy: In Vivo and In Vitro Studies

Jun Lu1ABCDEFG, Nina Zhou1BCDE, Ping Yang1BC, Lanqiuzi Deng1DEF, Ganzhe Liu2AEF*

DOI: 10.12659/MSM.916458

Med Sci Monit 2019; 25:8499-8508

Abstract

BACKGROUND: This study aimed to discover the effect and mechanism of microRNA-27a-3p (miR-27a-3p) in epilepsy.

MATERIAL AND METHODS: To perform our investigation, in vivo and in vitro models of epilepsy were induced using kainic acid (KA). Expression of miR-27a-3p in the hippocampus of epileptic rats or normal rats or neuronal cells was detected using quantitative reverse transcription polymerase chain reaction (qRT-PCR). Racine score was used to assess seizures in epileptic rats. Cell viability and cell apoptosis were analyzed by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay and flow cytometry. Enzyme-linked immunosorbent assay (ELISA) was performed to detect inflammatory factors expression.

RESULTS: Significantly higher expression of miR-27a-3p in the hippocampus of epileptic rats and in KA-induced neurons was observed. We found that miR-27a-3p inhibitor alleviated seizures in epileptic rats. miR-27a-3p inhibitor also inhibited apoptosis of hippocampal neurons in epileptic rats, promoted Bcl2 expression, and decreased Bax and Caspase3 expression. The results showed that miR-27a-3p inhibitor effectively reduced the expression levels of interleukin-1β (IL-1β), IL-6, and tumor necrosis factor-α (TNF-α) in hippocampal tissues of epileptic rats. Dual luciferase reporter assay showed that mitogen-activated protein kinase 4 (MAP2K4) was a direct target of miR-27a-3p. miR-27a-3p inhibitor significantly promoted the cell viability of KA-induced neurons, inhibited cell apoptosis, promoted the expression of Bcl-2, and decreased Bax and Caspase3 expression, and all these changes were abolished by MAP2K4-siRNA co-transfection.

CONCLUSIONS: Our preliminary findings indicated that miR-27a-3p inhibitor protected against epilepsy-induced inflammatory response and hippocampal neuronal apoptosis by targeting MAP2K4.

Keywords: Apoptosis, Epilepsy, MicroRNAs, Neurons, Cell Survival, Disease Models, Animal, Down-Regulation, Gene Expression Regulation, HEK293 Cells, Hippocampus, Inflammation, Kainic Acid, MAP Kinase Kinase 4, Male, RNA, Small Interfering, Rats, Rats, Sprague-Dawley, transcriptional activation, Tumor Necrosis Factor-alpha

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Medical Science Monitor eISSN: 1643-3750
Medical Science Monitor eISSN: 1643-3750