戊四氮点燃癫大鼠海马巢蛋白和骨形成蛋白-4的表达研究

Experimental study of Nestin and BMP4 expression in the hippocampus of pentylenetetrazol (PTZ) kindled epilepsy rats

目的观察巢蛋白(nestin)和骨形成蛋白4(BMP4)基因在戊四氮(PTZ)点燃癫大鼠海马中的表达,并探讨两者与癫发病机制的关系。方法将81只成年雄性SD大鼠随机分为实验组(n=54)和对照组(n=27)。实验组采用PTZ点燃癫大鼠,按点燃中的不同时相点,又随机分为9组。用免疫组化技术、地高辛标记特异性寡核苷酸探针原位杂交组织化学技术,观察海马nestin和BMP4表达的变化。结果nestin阳性细胞在PTZ注射后3d开始出现在齿状回、CA3区和CA1区,到7d达到高峰,以后逐渐减少。BMP4在PTZ注射后7d开始增多,在点燃后1d达到高峰,以后逐渐减少,主要分布在齿状回、CA3区和CA1区。结论PTZ点燃可引起海马内星形胶质细胞增生、活化和神经发生,这可能是癫海马组织胶质化、神经元可塑性的病理基础;BMP4可能在PTZ癫形成过程中起重要作用。

Objective To investigate the expression of nestin and BMP4 in different hippocampal areas of PTZ kindled epilepsy rats and explore their relationship with the pathogenesis of epilepsy. Methods 81 male SD rats were randomly divided into experimental group（ n =54） and control group（ n =27 ）. Rats of experimental group were man-made epilepsies models by chemical method based on the process of PTZ kindling. The experimental group was divided into 9 groups according to different time phase, lmmunohistochemistry, in situ hybridization labeled with Dig-oligonucleotide probe were applied to observe the expression of nestin and BMP4 in hippocampus. Results After infusion of PTZ, the nestin-pesitive cells in the experimental group appeared in dentate gyrus,CA3 ,CA1 on the third day and reached maximal expression level on the seventh day and then decreased gradually. In the PTZ kindled epilepsy rats, BMP4-pesitive cells were mainly located in dentate gyrus, CA3, CA1 with the number increasng on seventh day after PTZ infision, and reaching maximal expression level on the next day of kindling, then decreasing gradually. Conclusions PTZ kindling can induce proliferation, activation and neurogenesis of astrocytes in hippoeampus, which may serve as the pathological base of scarring and neuronal plasticity of the hippocampus epileptic tissue; BMP4 may play a critical role in the pathogenesis of PTZ kindling epilepsy.