癫痫发作1h后延髓内脏带区域内反应神经元与星形胶质细胞功能单位的分布特征:激光共聚焦显微镜研究

背景传统观点认为癫痫是大脑不同区域内神经元出现异常兴奋引起的复杂神经行为紊乱现象.而对星形胶质细胞在癫痫发病中的作用目前研究较少.目的研究戊四氮诱导大鼠癫痫发作后延髓内脏带内神经元和星形胶质细胞的反应.设计随机对照的实验研究.地点和材料实验在南方医科大学珠江医院神经外科实验室和解放军第四军医大学全军神经科学研究所完成.成年健康SD大鼠14只,体质量180～220 g,清洁级,由解放军第四军医大学实验动物中心提供.干预用抗Fos蛋白、抗酪氨酸羟化酶和抗胶质原纤维酸性蛋白的三重免疫荧光组织化学方法结合激光共聚焦显微镜技术,显示癫痫发作1 h后延髓内脏带内反应性神经元与星形胶质细胞的分布.主要观察指标对延髓内脏带内Fos,胶质原纤维酸性蛋白和抗酪氨酸羟化酶阳性细胞的分布及胶质原纤维酸性蛋白阳性星形胶质细胞与神经元的关系进行观察.结果延髓内脏带内的Fos阳性神经元和胶质原纤维酸性蛋白阳性星形胶质细胞明显增多,三重免疫组化方法显示反应性神经元(Fos阳性)与反应性星形胶质细胞(胶质原纤维酸性蛋白阳性)关系密切,发现3种不同标记的"神经元-星形胶质细胞复合体"即抗酪氨酸羟化酶+/Fos+/胶质原纤维酸性蛋白+三标记复合体、抗酪氨酸羟化酶+/胶质原纤维酸性蛋白+/Fos-和Fos+/胶质原纤维酸性蛋白+/抗酪氨酸羟化酶-二标记复合体.结论延髓内脏带内的神经元和星形胶质细胞在癫痫发作时反应强烈,可能以神经元-星形胶质细胞复合体作为功能单位参与癫痫发病的调节.

Distribution character of functional unit of reaction neuron and astrocyte in medullary visceral zone 1 hour after epileptic attack: a laser confocal microscopic research

BACKGROUND: It is considered traditionally that epilepsy is a kind of complicated nervous conduct disorder caused by abnormally excited neuron in different area in brain. While the research on the function of astrocyte in epileptic attack is very rare.OBJECTIVE: To study the reaction of neuron and astrocyte in medullary visceral zone after epilepsy induced by pentetrazole in rats.DESIGN: A randomized controlled experimental research.SETTING and MATERIALS: The experiment was done in the Neurosurgery Laboratory of Zhujiang Hospital Affiliated to Southern Medical University and Neuroscience Institute of Fourth Military Medical University of Chinese PLA. Fourteen healthy adult SD rats, weighing 180 - 220 g, clean grade, were provided by the Experimental Animal Center of Fourth Military Medical University of Chinese PLA.INTERVENTIONS: Distribution of neuron and astrocyte in MVZ 1 hour after epileptic attack was shown by laserconfocal microscopic technique combined with triplication immunofluorescence histochemistry of anti-Fos protein, anti-tyrosine hydroxylase(TH) and anti-glial fibrillary acidic protein(GFAP).MAIN OUTCOME MEASURES: Observation of distribution of positive cells of Fos, GFAP and TH in MVZ and relationship between GFAP positive astrocyte and neuron.RESULTS: Fos positive neurons and GFAP positive astrocytes in MVZ increased significantly. Triplication immunofluorescence histochemistry showed reaction neuron(Fos positive) closely related with reaction astrocyte(GFAP positive) . Three kinds of N-ASC compounds with different labels were found, which were TH +/Fos +/GFAP + three labeled compound, TH + /GFAP +/Fos- and Fos+/GFAP +/TH- two labeled compound.CONCLUSION: Neuron and astrocyte in MVZ reacted strongly when epilepsy attacks. N-ASC as a functional unit may regulate onset of epilepsy.