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Mitochondrial damage in hippocampal neurons of rats with epileptic protein expression of Fas and caspase-3

机译:Fas和caspase-3癫​​痫蛋白表达对大鼠海马神经线粒体的损​​伤

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摘要

Epilepsy model in rats was established to observe the behavior and pathological changes, and to detect mitochondrial dysfunction, exploring its possible molecular mechanisms. The epileptic status of Sprague-Dawley (SD) rats was induced by intraperitoneal injection of lithium chloride, and the change of behavior was recorded. Electroencephalogram (EEG) was used to measure the abnormal discharge of neurons in rats. The brain tissue was fixed with polyformaldehyde and the paraffin sections were prepared, and the damage of the hippocampal neurons was observed with Nissl staining. Mitochondrial ATP and mitochondrial DNA were examined to assess mitochondrial dysfunction. Finally, qPCR and western blot analysis were used to detect mRNA and protein expression of fatty acid synthetase (Fas), Fas ligand (FasL) and caspase-3 in rat hippocampal neurons. The correlation between the mitochondrial dysfunction of rat hippocampal neurons and Fas and caspase-3 was analyzed. Compared with the normal group rats, the model group showed typical seizures, which were determined by the Racine attack score. EEG of the hippocampus of the model group was recorded in cluster in model group rats. Nissl staining showed a different degree of damage to the hippocampal neurons in the model group compared with normal rats. The mitochondrial ATP content and DNA content of rat hippocampal neurons in the model group were significantly lower than that of normal rats (P<0.01). The qPCR and western blot results showed that the mRNA and protein expression levels of Fas, FasL and caspase-3 were significantly increased in the hippocampus of rat model group (P<0.01). The expression level of Fas and caspase-3 in hippocampal tissues of rats was negatively correlated with mitochondrial DNA content. In conclusion, seizures cause damage of neuron mitochondria in rat hippocampus leading to death of hippocampal neurons, the mitochondrial damage of hippocampal neurons in epileptic rats was closely related to the expression of Fas and caspase-3.
机译:建立大鼠癫痫模型,观察其行为和病理变化,并检测线粒体功能障碍,探讨其可能的分子机制。腹膜内注射氯化锂诱导Sprague-Dawley(SD)大鼠的癫痫状态,并记录其行为变化。脑电图(EEG)用于测量大鼠神经元的异常放电。用聚甲醛固定脑组织,制备石蜡切片,用尼氏染色观察海马神经元的损伤。检查线粒体ATP和线粒体DNA以评估线粒体功能障碍。最后,用qPCR和western blot分析检测大鼠海马神经元中脂肪酸合成酶(Fas),Fas配体(FasL)和caspase-3的mRNA和蛋白表达。分析了大鼠海马神经元线粒体功能障碍与Fas和caspase-3的相关性。与正常组大鼠相比,模型组表现出典型的癫痫发作,这由拉辛发作评分决定。在模型组大鼠中成簇记录模型组海马的EEG。与正常大鼠相比,Nissl染色显示模型组对海马神经元的损伤程度不同。模型组大鼠海马神经元线粒体ATP含量和DNA含量均明显低于正常大鼠(P <0.01)。 qPCR和Western blot结果表明,模型组海马中Fas,FasL和caspase-3的mRNA和蛋白表达水平显着升高(P <0.01)。大鼠海马组织中Fas和caspase-3的表达水平与线粒体DNA含量呈负相关。总之,癫痫发作引起大鼠海马神经元线粒体损伤,导致海马神经元死亡,癫痫大鼠海马神经元线粒体损伤与Fas和caspase-3的表达密切相关。

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