Expression and significance of signaling pathway of miR-134/CREB/pCREB in patients with epilepsy and in epileptic rats
Received:September 19, 2014  
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KeyWord:miR-134;CREB;pCREB;Refractory epilepsy;Synaptic plasticity
王倩 重庆医科大学附属第二医院神经内科, 重庆
陈阳美 重庆医科大学附属第二医院神经内科, 重庆
郭靖 重庆医科大学附属第二医院神经内科, 重庆
杨小兰 资阳市乐至县人民医院神经内科, 四川 资阳
谢运兰 重庆医科大学附属第二医院中心实验室, 重庆
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      Objective To investigate the expression of microRNA-134 (miR-134), CREB and pCREB in the temporal lobe tissue of patients and epileptic rats and to explore their roles in pathogenesis of epilepsy. Methods Temporal lobe tissue samples of 14 patients with refractory epilepsy and 10 non-epileptic patients, and hippocampus and brain tissue samples of 42 rats were used in this study. Forty-two healthy adult male Sprague-Dawley rats were randomly divided into 6 epilepsy groups (24 h, 72 h, 7 d, 14 d, 30 d, and 60 d after kindling epilepsy) and a normal control group (n=6 for all groups). The rat model of epilepsy was generated by intraperitoneal injection of 127 mg/kg lithium chloride and 16-20 h later, 35 mg/kg pilocarpine. In the temporal lobe tissue of patients and hippocampal tissue of rats, the expression level of miR-134 was detected by real-time polymerase chain reaction. The expression levels of CREB and pCREB were determined by Western blot, and CREB and pCREB localization was assessed by immunohistochemistry. Results Compared with the control rats, the expression of miR-134 was significantly decreased in the temporal lobe tissue of experimental rats at 72 h,7 d,14 d, 60 d after kindling (P<0.05),and no significant change at 24 h and 30 d after kindling (P>0.05). Expression of miR-134 in patients with refractory epilepsy was significantly lower than that of the controls (P<0.05), while up-regulation of CREB expression was at the same time points (P<0.05). Up-regulation of pCREB expression was at all the time points after kindling (P<0.05). CREB and p-CREB expressions were seen in the nuclei of neurons, and significantly higher in patients with refractory epilepsy and epileptic rats. Conclusions The expression of miR-134 is significantly decreased and that of CREB and pCREB was significantly increased in the temporal lobe tissue of patients with refractory epilepsy and the hippocampal tissue of epileptic rats. These findings indicate that the signaling pathway of miR-134/CREB/pCREB may play an important role in the pathogenesis of epilepsy.
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