The Protective Effect Of Amentoflavone On Hippocampus Neurons In Epileptic Mice

Objective Research the effect of amentoflavone on seizures and hippocampalneurons in the model of kindling mice induced by pilocarpine, and to explore themechanism.Methods A kindling model of pilocarpine induced epilepsy in mice wasestablished, and intervened with amentoflavone. After modelled, apply withneuroelectrophysiology, pathology and molecular biology technology, to analysisthe influence of amentoflavone on kindling mice and the protective effect onhippocampal neurons.Results1. Ethology manifestations and EEG: Blank mice without seizures. After theinjection of pilocarpine, compared with Blank group，mice of behavioral and EEGin AT group, EP group was instability, showing persistent epileptiform discharges,animal accompanied Ⅲ~Ⅴ grade seizures; AP group and VPA group mice have nogrand mal seizures occurred, occasionally I~Ⅱ grade seizures, and seizures wassignificantly shorter, EEG manifestations were similar to the Blank group.2. Nissl staining: The number of hippocampal CA1neurons in AP group(121.40±8.72) and Blank group (125.20±9.96) without significantly reduced,neuronal sharp edges, arranged in neat rows, visible in the cytoplasm Nissl bodies.The neurons in AT group (84.60±6.19), VPA group (70.25±4.34) and the EPgroup (65.30±7.53) were in a significant absence state, neurons are arrangedloosely, a serious lack of cytoplasmic staining shades nucleolus disappeared or not obvious.3. Immunohistochemistry: Mean optical density value method was used todetect the expression of NF-κB p65. The expression of AP Group, AT group, VPAgroup, EP group and Blank groups NF-κB p65were4204.26±116.30,15291.21±501.56,35599.58±2742.78,35710.87±2570.31,3185.12±251.35, respectively.Compared with the EP group, AT group, VPA group, the expression levels ofNF-κB p65in AP group and Blank group were statistically significant difference(P <0.01).4. TUNEL staining: In the amentoflavone pre-treated, neurons were fairlywell preserved and sparse damage was observed in CA1subfield. Most of themorphologically damaged neurons stained with cresyl violet were also positive forTUNEL. Sparse TUNEL-positive cells were observed in CA1subfield in the APgroup (1.70±0.98vs. the EP group value of12.55±3.25, P <0.05). No significantdifference was observed between the Blank group (0.55±0.51) and AP group.5. Western Blot: There was no significant difference with the expression ofNF-κB p65protein between AP group (0.49±0.13) and Blank group (0.33±0.12).Compared with Blank group， the expression of NF-κB p65protein in AT group(0.71±0.09), VPA group (1.18±0.19) and EP group (1.10±0.17) weresignificantly increased (P <0.01).Conclusion Amentoflavone exerts a significant preventive effect on seizuresand neuroprotection in hippocampus, and its mechanisms related to the inhibitoryeffect of amentoflavone on inflammatory response of brain in epilepsy process.