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目的探讨遗传性癫痫大鼠(TRM)海马内GAT-1和GAT-3mRNA与蛋白的表达。方法应用RT-PCR技术检测GAT-1和GAT-3mRNA的表达;应用Westernblot方法检测GAT-1和GAT-3蛋白的表达;应用免疫组化方法定位GAT-1和GAT-3蛋白的分布。结果TRM大鼠海马内GAT-1mR-NA和蛋白表达均高于正常Wistar大鼠(P<0.01);GAT-3mRNA表达高于正常Wistar大鼠(P<0.05);GAT-3蛋白表达亦高于正常Wistar大鼠(P<0.01);免疫组化结果显示GAT-1在TRM和Wistar大鼠海马的CA1、CA3和DG区均有明显分布,且在TRM鼠海马的CA1、CA3和DG区中均比在正常Wistar鼠中明显;而GAT-3蛋白只在TRM和Wistar大鼠海马的CA1区中表达丰富,同时,和正常Wistar鼠比较,TRM鼠海马CA1区中GAT-3蛋白的表达明显增加。结论TRM大鼠海马内GAT-1和GAT-3表达上调可能促进了癫痫的发生,可能是癫痫治疗的靶点。
Objective To investigate the expression of GAT-1 and GAT-3 mRNA and protein in the hippocampus of hereditary epilepsy rats (TRM). Methods The expression of GAT-1 and GAT-3 mRNA was detected by RT-PCR. The expression of GAT-1 and GAT-3 protein was detected by Western blot. The distribution of GAT-1 and GAT-3 protein was detected by immunohistochemistry. Results The expression of GAT-1mR-NA and protein in hippocampus of TRM rats were higher than those in normal Wistar rats (P <0.01). The expression of GAT-3 mRNA in hippocampus of TRM rats was higher than that in normal Wistar rats (P <0.05) In normal Wistar rats (P <0.01), immunohistochemistry results showed that GAT-1 was obviously distributed in CA1, CA3 and DG of hippocampus of TRM and Wistar rats, and in CA1, CA3 and DG of hippocampus of TRM rats , While GAT-3 protein was only abundantly expressed in CA1 region of hippocampus of TRM and Wistar rats. Meanwhile, compared with normal Wistar rats, GAT-3 protein expression in hippocampal CA1 region of TRM mice obviously increase. Conclusion The up-regulation of GAT-1 and GAT-3 in hippocampus of TRM rats may promote the occurrence of epilepsy and may be the target of epilepsy treatment.