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目的探讨托吡酯对TRM大鼠脑内GAT-1和GAT-3 mRNA和蛋白表达的影响。方法分别应用实时定量PCR法和Western blot方法检测Wistar正常组、TRM组、TPM40组和TPM80组脑内GAT-1和GAT-3 mRNA和蛋白表达。结果 GAT-1、GAT-3和β-actin标准曲线相关性良好,基因特异性强。GAT-1 mRNA在TRM组的海马高于正常组(P<0.01);与TRM组比较,TPM80组海马中GAT-1 mRNA明显降低(P<0.05);GAT-3 mRNA在TRM组海马中的表达高于正常组(P<0.05);GAT-1蛋白表达在TRM组的海马高于正常组(P<0.01);与TRM组比较,TPM80组海马中GAT-1蛋白明显降低(P<0.01);GAT-3蛋白在TRM组海马中的表达高于正常组(P<0.05)。结论 TRM组海马内GAT-1及GAT-3的表达增加可能促进了癫痫的发生。托吡酯的抗癫痫作用可能是通过降低海马内GAT-1的表达来实现的,这种调节存在剂量依赖性。
Objective To investigate the effects of topiramate on GAT-1 and GAT-3 mRNA and protein expression in the brain of TRM rats. Methods The expressions of GAT-1 and GAT-3 mRNA and protein in brain tissues of Wistar normal group, TRM group, TPM40 group and TPM80 group were detected by real-time quantitative PCR and Western blot respectively. Results GAT-1, GAT-3 and β-actin standard curve of good correlation, gene-specific strong. GAT-1 mRNA in hippocampus of TRM group was higher than that in normal group (P <0.01). Compared with TRM group, GAT-1 mRNA in hippocampus of TPM80 group was significantly lower (P <0.05); GAT- The expression of GAT-1 protein in hippocampus of TRM group was higher than that in normal group (P <0.01). Compared with TRM group, GAT-1 protein in hippocampus of TPM80 group was significantly decreased (P <0.01) ). The expression of GAT-3 protein in hippocampus of TRM group was higher than that in normal group (P <0.05). Conclusion The increased expression of GAT-1 and GAT-3 in the hippocampus of TRM group may promote the occurrence of epilepsy. The antiepileptic effect of topiramate may be achieved by decreasing the expression of GAT-1 in the hippocampus, which is dose-dependent.