Amentoflavone Affects Epileptogenesis and Exerts Neuroprotective Effects by Inhibiting NLRP3 Inflammasome.

Shikuo Rong,Shenhai Liu,Peng Zhang,Xinxiao Li,Kuisheng Sun,Yayun Fan,Xiaoliang Xie,Feng Wang,Tao Sun,Ding Wan,Junming Huo

doi:10.3389/fphar.2019.00856

Abstract

Brain inflammation is one of the main causes of epileptogenesis, a chronic process triggered by various insults, including genetic or acquired factors that enhance susceptibility to seizures. Amentoflavone, a naturally occurring biflavonoid compound that has anti-inflammatory effects, exerts neuroprotective effects against nervous system diseases. In the present study, we aimed to investigate the effects of amentoflavone on epilepsy in vivo and in vitro and elucidate the underlying mechanism. The chronic epilepsy model and BV2 microglial cellular inflammation model were established by pentylenetetrazole (PTZ) kindling or lipopolysaccharide (LPS) stimulation. Cognitive dysfunction was tested by Morris water maze while hippocampal neuronal apoptosis was evaluated by immunofluorescence staining. The levels of nucleotide oligomerization domain-like receptor protein 3 (NLRP3) inflammasome complexes and inflammatory cytokines were determined using quantitative real-time polymerase chain reaction, Western blotting, immunofluorescence staining, and enzyme-linked immunosorbent assay. Amentoflavone reduced seizure susceptibility, minimized PTZ-induced cognitive dysfunction, and blocked the apoptosis of hippocampal neurons in PTZ-induced kindling mice. Amentoflavone also inhibited the activation of the NLRP3 inflammasome and decreased the levels of inflammatory cytokines in the hippocampus of PTZ-induced kindling mice. Additionally, amentoflavone could alleviate the LPS-induced inflammatory response by inhibiting the NLRP3 inflammasome in LPS-induced BV2 microglial cells. Our results indicated that amentoflavone affects epileptogenesis and exerts neuroprotective effects by inhibiting the NLRP3 inflammasome and, thus, mediating the inflammatory process in PTZ-induced kindling mice and LPS-induced BV2 microglial cells. Therefore, amentoflavone may be a potential treatment option for epilepsy.

Highlights

Epilepsy, a common chronic brain disorder and the most common severe neurological condition (Loscher et al, 2013; Lukawski et al, 2016), affects ~1% of the population and all ages
These results suggest that amentoflavone reduces seizure susceptibility and exerts antiepileptic seizure activity in PTZ kindled mice
We revealed that amentoflavone reduced seizure susceptibility, minimized cognitive dysfunction, and blocked the apoptosis of hippocampal neurons in PTZ-induced kindling mice

Summary

Introduction

A common chronic brain disorder and the most common severe neurological condition (Loscher et al, 2013; Lukawski et al, 2016), affects ~1% of the population and all ages. Epilepsy often requires lifelong medication (Hauser et al, 2018), and as more than 30% of patients suffering from this disorder do not achieve good control with available drugs, this condition often progresses to drug-resistant epilepsy (Laxer et al, 2014). Developing new drugs to prevent or treat epilepsy is still urgently needed. Anti-inflammatory therapies may serve as an intervention and could be a promising strategy for preventing and treating seizures and its related neurobehavioral comorbidities

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