Amentoflavone Affects Epileptogenesis and Exerts Neuroprotective Effects by Inhibiting NLRP3 Inflammasome

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.

Connexins-Based Hemichannels/Channels and Their Relationship with Inflammation, Seizures and Epilepsy

“…The assembly of the NLRP3 inflammasome results in the conversion of procaspase-1 from its pro form to its active form, which then processes the cleavage of several substrates, such as pro-IL-1β and pro-IL-18, in the mature cytokines IL-1β and IL-18 [108]; this process can aggravate inflammation. In addition, numerous studies have implicated neuroinflammation as the cause, and result, of epilepsy [109,110].…”

Inhibitory Activity of Fruits Extracts of <i>A</i><i>n</i><i>t</i><i>i</i><i>d</i><i>e</i><i>s</i><i>m</i><i>a </i><i>b</i><i>unius</i><i> </i>on the Proliferation and Migration of MDA-MB-231 Breast Cancer Cells

“…The flavonoids in the fruit extract of A. bunius such as coumarin, genistin, amentoflavone and luteolin 7-galactoside have already been proved to have multiple biological activities [15,16,17]. Amentoflavone is a naturally occurring biflavonoid compound abundant in Selaginella tamariscina, with antiinflammatory [18], anti-oxidative [19], anti-microbial [20], anti-apoptotic [21], anti-radiation [22], antitumor [12], neuroprotective [23,24] and cyano-bacterial killing effects [25]. Previously, the authors confirmed the antiproliferative effect of the ethanolic extract of leaves of Antidesma montanum on the MDA-MB-231 cells and found that amentoflavone was present in this extract [26].…”

Inhibition of Death-associated Protein Kinase 1 protects against Epileptic Seizures in mice

“…Stage 0: normal behavior; stage 1: immobility; stage 2: facial and neck jerks; stage 3: tail extension; stage 4: chronic seizure (sitting); stage 5: generalized chronic, tonic-clonic seizure (lying on belly); stage 6: generalized chronic, tonic-clonic seizure (lying on side); stage 7: full tonic extension; and stage 8: death [35][36][37]. Kindling paradigm: mice were treated with a subconvulsive dose of PTZ (35 mg/kg) every other day, observed for the next 15 min and scored for seizure severity after each PTZ injection based on a modified Racine scale [38][39][40]. Stage 0: normal behavior; stage 1: immobilization or facial myoclonus; stage 2: head nodding; stage 3: forelimb clonus or short myoclonic jerks; stage 4: rearing generalized chronic seizure; stage 5: generalized tonic-clonic seizures and falling with loss of posture; and stage 6: death.…”