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Inhibition of microRNA-34a Suppresses Epileptiform Discharges Through Regulating Notch Signaling and Apoptosis in Cultured Hippocampal Neurons

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Abstract

Epilepsy is characterized by recurrent unprovoked seizures and some seizures can cause neuronal apoptosis, which is possible to make contributions to the epilepsy phenotype, impairments in cognitive function or even epileptogenesis. Moreover, many studies have indicated that microRNA-34a (miRNA-34a) is involved in apoptosis through regulating Notch signaling. However, whether miRNA-34a participates in neuronal apoptosis after seizures remain unclear. Therefore, we aimed to explore the expression of miRNA-34a and its effects on the epileptiform discharge in spontaneous recurrent epileptiform discharges (SREDs) rat hippocampal neuronal pattern. Mg2+-free medium was used to induce SREDs, quantitative reverse-transcription polymerase chain reaction was used to detect the expression of miRNA-34a, western blot was used to determine the expression of Notch pathway and apoptosis-related proteins, and whole cell current clamp recordings was used to observe the alteration of epileptiform discharge. We found obvious apoptosis, increased expression of miRNA-34a and decreased expression of Notch signaling in Mg2+-free-treated neurons. Treatment with miRNA-34a inhibitor decreased the frequency of action potentials, activated Notch signaling and prevented neuronal apoptosis in Mg2+-free-treated neurons. However, treatment with miRNA-34a mimics increased the frequency of action potentials, down-regulated Notch signaling and promoted neuronal apoptosis in Mg2+-free-treated neurons. Furthermore, γ-secretase inhibitor N-[N-(3,5-di-uorophenacetyl)-1-alanyl]-S-phenylglycine t-butylester (DAPT), an inhibitor of Notch signaling, could weaken anti-apoptosis effect of miRNA-34a inhibitor. These results suggest that inhibition of miRNA-34a could suppress epileptiform discharges through regulating Notch signaling and apoptosis in the rat hippocampal neuronal model of SREDs.

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Abbreviations

SREDs:

Spontaneous recurrent epileptiform discharges

qRT-PCR:

Quantitative reverse-transcription polymerase chain reaction

MAP2:

Protein-2 associated with anti-microtubule

pBRS:

Physiological basal recording solution

AP:

Action potentials

Bax:

Bcl-2-associated X protein

Bcl-2:

B-cell lymphoma-2

HRP:

Horseradish peroxidase

NIC:

Notch intracellular domain

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Authors and Affiliations

  1. Department of Pediatrics, Jinan Central Hospital, Shandong University, Number 105, Jiefang Road, Lixia District, Jinan, 250012, Shandong, China

    Jinli Wang, Yuan Zheng, Fenfen Xu, Xiao Zhou & Hongyang Zhao

  2. Clinical Medical College, Taishan Medical University, Taian, Shandong, China

    Xu Cheng & Piaopiao Zhang

Authors
  1. Jinli Wang
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  2. Yuan Zheng
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  3. Xu Cheng
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  4. Fenfen Xu
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  5. Piaopiao Zhang
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  6. Xiao Zhou
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  7. Hongyang Zhao
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Correspondence to Hongyang Zhao.

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Every animal use process was supported by the Animal Care and Use Committee of Shandong University and was in severe accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals.

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Wang, J., Zheng, Y., Cheng, X. et al. Inhibition of microRNA-34a Suppresses Epileptiform Discharges Through Regulating Notch Signaling and Apoptosis in Cultured Hippocampal Neurons. Neurochem Res 44, 1252–1261 (2019). https://doi.org/10.1007/s11064-019-02772-x

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  • DOI: https://doi.org/10.1007/s11064-019-02772-x

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