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目的了解小清蛋白(PV)、钙视网膜结合蛋白(CR)在皮质发育不良(CD)大鼠大脑皮质中的表达,探讨其在鉴别CD和参与癫痫发病机制中的作用。方法孕15d的SD大鼠腹腔注射卡莫司汀建立子代鼠CD模型,P30行热水浴诱发惊厥实验,组织病理学技术观察仔鼠皮质结构改变,免疫组化、免疫荧光、RT-PCR技术检测PV、CR在大鼠大脑皮质的表达。结果①模型鼠热水浴诱发惊厥潜伏期缩短;②免疫组化示模型鼠大脑皮质PV阳性细胞数量([77.50±4.49个)/视野]较对照组([37.28±2.72)个/视野]减少(P<0.05)、空间分布紊乱、皮质浅层出现阳性细胞结节和阳性细胞空白区,CR的表达较对照组无显著差异;③免疫荧光示模型鼠PV阳性细胞减少,胞体增大;④RT-PCR检测示模型鼠PVmRNA相对表达量显著降低,CRmRNA的相对表达量无显著差异。结论PV可作为鉴定CD的指标;表达PV的GABA能抑制性中间神经元的损伤可能在CD所致的难治性癫痫中具有重要作用。
Objective To investigate the expression of small albumin (PV) and calcium retinal binding protein (CR) in the cerebral cortex of cortical dysplasia (CD) rats, and to explore the role of PV and CD in the pathogenesis of epilepsy. Methods The SD rats were injected intraperitoneally with carmustine to establish the CD model of offspring mice. P30 was induced by hot bath in convulsion. Histopathology was used to observe the changes of cortical structure, immunohistochemistry, immunofluorescence and RT-PCR The technique was used to detect the expression of PV and CR in rat cerebral cortex. Results ① The latent period of convulsion was shortened in hot water bath induced by model rats; ② The number of PV-positive cells in cerebral cortex ([77.50 ± 4.49 / visual field] in model group was decreased compared with that in control group ([37.28 ± 2.72] / visual field] P <0.05), the spatial distribution of disorders, cortical superficial layer of positive cells nodules and positive cells in the blank area, the expression of CR was no significant difference compared with the control group; ③ immunofluorescence model mice PV positive cells decreased, cell body increased; ④ RT- PCR showed that the relative expression level of PVmRNA in model mice was significantly decreased, while the relative expression level of CRmRNA was not significantly different. Conclusions PV can be used as an index to identify CD. GABA-inhibitory interneurons that express PV may play an important role in refractory epilepsy induced by CD.