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MiR-139-5p Upregulation Alleviated Spontaneous Recurrent Epileptiform Discharge-induced Oxidative Stress and Apoptosis in Rat Hippocampal Neurons via Regulating the Notch Pathway.

Abstract：

:Epilepsy was characterized by the occurrence of spontaneous recurrent epileptiform discharges (SREDs) in neurons. Previous studies suggested that microRNA (miR)-139-5p and the Notch pathway were implicated in Epilepsy; however, their interaction remained vague. Rat primary hippocampal neurons were isolated and identified by immunofluorescence staining. The cells were then used for SREDs model construction and further subjected to flow cytometry for apoptosis detection. Contents of lactate dehydrogenase (LDH), malondialdehyde (MDA), super oxidase dismutase (SOD) contents, and reactive oxygen species (ROS) and the level of mitochondrial membrane potential (MMP) were determined using commercial kits. Target gene and potential binding sites of miR-139-5p were predicted with TargetScan and confirmed by dual-luciferase reporter assay. Expressions of miR-139-5p, Notch pathway-related proteins and apoptosis-related proteins were measured by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot as needed. The results showed that the hippocampal neurons were microtubule-associated protein 2 (MAP2)-positive. MiR-139-5p was downregulated in SREDs model cells. SREDs promoted apoptosis and increased the contents of LDH, MDA, and ROS and the level of MMP while reducing miR-139-5p expression and SOD content in cells, which was reversed by miR-139-5p overexpression. Notch-1 was recognized as the target gene of miR-139-5p, and its expression was negatively regulated by miR-139-5p. Besides, Notch-1 overexpression reversed the effects of miR-139-5p upregulation on the expressions of Notch pathway-related proteins and apoptosis-related proteins, cell apoptosis, oxidative stress and MMP in SREDs-treated cells. Our results indicated that miR-139-5p upregulation alleviated SREDs-induced oxidative stress and cell apoptosis via regulating the Notch pathway, which provides new insights into the role of miRNA in the occurrence and development of epilepsy. This article is protected by copyright. All rights reserved.

journal_name

Cell Biol Int

journal_title

Cell biology international

authors

Zhao C,Yang F,Wei X,Zhang J

doi

10.1002/cbin.11509

subject

Has Abstract

pub_date

2020-11-28 00:00:00

eissn

1065-6995

issn

1095-8355

pub_type

杂志文章

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MiR-139-5p Upregulation Alleviated Spontaneous Recurrent Epileptiform Discharge-induced Oxidative Stress and Apoptosis in Rat Hippocampal Neurons via Regulating the Notch Pathway.