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酸枣仁皂苷B对急性癫痫发作的缓解及神经保护作用

Anticonvulsant and neuroprotective effects of jujuboside B on acute epilepsy

  • 摘要:
    目的  探讨酸枣仁皂苷B(jujuboside B, JuB)对小鼠急性癫痫发作的缓解及海马神经元保护作用。
    方法  使用莰烯氯噻嗪(cyclothiazide, CTZ)刺激原代海马神经元,构建体外癫痫模型。使用膜片钳技术记录神经元自发癫痫样放电。预先向成年雄性C57BL/6小鼠腹腔注射JuB,利用戊四唑(pentylenetetrazol, PTZ)诱导小鼠急性癫痫模型,记录癫痫发作严重程度。造模24 h后,处死小鼠并取材。使用蛋白质印迹法检测海马内线粒体凋亡相关蛋白的表达量,病理学染色观察海马CA1区和CA3区神经元损伤。
    结果 JuB可以降低CTZ诱导的癫痫样放电频率(P<0.001);30 mg/kg和50 mg/kg JuB预处理可以降低PTZ诱导的小鼠癫痫发作最高行为学评分,并延长Racine Ⅲ级发作潜伏期(P<0.001)。与模型组相比,JuB可以降低急性癫痫小鼠Bax蛋白表达(P<0.01),提高Bcl-2蛋白表达(P<0.05),保护小鼠海马CA1区(P<0.05)和CA3区(P<0.01)神经元活性,改善细胞排列紊乱、边界不清、核固缩碎裂溶解等病理形态。
    结论 JuB在体内和体外均表现出癫痫抑制作用,并可以抑制急性癫痫模型中线粒体凋亡、减轻海马神经元损伤,具有潜在抗癫痫作用。

     

    Abstract:
    Objective  To investigate the alleviating effect of jujuboside B (JuB) on acute epilepsy in mice and its protective effect on hippocampal neurons.
    Methods An in vitro epilepsy model was established by stimulating primary hippocampal neurons with cyclothiazide (CTZ). Spontaneous epileptiform discharge was recorded using patch clamp technique. An acute epilepsy model was induced in adult male C57BL/6 mice by intraperitoneal injecting pentylenetetrazol (PTZ) following pretreatment with JuB, and severity of epilepsy was recorded. Mice were euthanized and brain tissues were collected 24 hours after model establishment. The expression levels of mitoptosis related proteins in the hippocampus were detected by Western blotting. Neuronal damage in the hippocampal CA1 and CA3 regions was observed using pathological staining.
    Results JuB reduced the frequency of CTZ-induced epileptiform discharges (P<0.001). Pretreatment with 30 mg/kg and 50 mg/kg JuB decreased the maximal behavioral seizure score and prolonged the latency to Racine stage Ⅲ seizures in PTZ-induced epileptic mice (P<0.001). Compared with the PTZ group, JuB treatment downregulated Bax protein level (P<0.01) and upregulated Bcl-2 protein level (P<0.05) in acute epileptic mice. Furthermore, JuB protected neuronal viability in the hippocampal CA1 (P<0.05) and CA3 (P<0.01) regions, and ameliorated pathological morphological changes including cellular disarray, unclear boundaries, pyknosis, fragmentation, and dissolution.
    Conclusions  JuB exhibits antiepileptic effects both in vivo and in vitro. It exerts potential antiepileptic effects by inhibiting mitoptosis and attenuating neuronal damage in the hippocampus in an acute epilepsy model.

     

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