Abstract：
BACKGROUND: Drug-resistant epilepsy (DRE) is a refractory neurological disorder. There is ample evidence that suggest that γ-aminobutyric acid-a (GABAA) receptors could be one of the mechanisms responsible for the development of drug resistance in epilepsy. It is also known that the cAMP response element binding protein (CREB) plays a possible key role in the transcriptional regulation of GABAA.
OBJECTIVE: This study explores the role of CREB in the development of DRE and the effect of CREB on GABA-related receptors in DRE.
METHODS: The CREB expression was increased or decreased in the hippocampus of normal rats by lentiviral transfection, who then underwent the lithium-pilocarpine-induced epilepsy model. Phenobarbital (PB) sodium and carbamazepine (CBZ) were used to select a drug-resistant epileptic model. The expression levels of GABAA receptor α1, β2, and γ2 subunits and CREB protein were measured in the rat hippocampus by western blot and fluorescent quantitative PCR.
RESULTS: The frequency and duration of seizures increased in the overexpression group compared to that in the control group. In addition, the severity, frequency, and duration of seizures decreased in the group with decreased expression. The hippocampus analysis of the expression levels of the CREB protein and CREB mRNA yielded similar findings. Altering the CREB protein expression in the rat hippocampus could negatively regulate the expression and transcript levels of GABAA receptors α1, β2, and γ2, suggesting that CREB may serve as a potential target for the development of treatment protocols and drugs for epilepsy.
CONCLUSIONS: Our study shows that enhanced CREB expression promotes the development of DRE and negatively regulates GABAA receptor levels and that the inhibition of CREB expression may reduce the incidence of DRE.
OBJECTIVE: This study explores the role of CREB in the development of DRE and the effect of CREB on GABA-related receptors in DRE.
METHODS: The CREB expression was increased or decreased in the hippocampus of normal rats by lentiviral transfection, who then underwent the lithium-pilocarpine-induced epilepsy model. Phenobarbital (PB) sodium and carbamazepine (CBZ) were used to select a drug-resistant epileptic model. The expression levels of GABAA receptor α1, β2, and γ2 subunits and CREB protein were measured in the rat hippocampus by western blot and fluorescent quantitative PCR.
RESULTS: The frequency and duration of seizures increased in the overexpression group compared to that in the control group. In addition, the severity, frequency, and duration of seizures decreased in the group with decreased expression. The hippocampus analysis of the expression levels of the CREB protein and CREB mRNA yielded similar findings. Altering the CREB protein expression in the rat hippocampus could negatively regulate the expression and transcript levels of GABAA receptors α1, β2, and γ2, suggesting that CREB may serve as a potential target for the development of treatment protocols and drugs for epilepsy.
CONCLUSIONS: Our study shows that enhanced CREB expression promotes the development of DRE and negatively regulates GABAA receptor levels and that the inhibition of CREB expression may reduce the incidence of DRE.
摘要：
背景：耐药性癫痫（DRE）是一种难治性神经系统疾病。有充分的证据表明，γ-氨基丁酸-a（GABAA）受体可能是导致癫痫耐药性发展的机制之一。还已知cAMP反应元件结合蛋白(CREB)在GABAA的转录调节中起可能的关键作用。
目的：本研究探讨了CREB在DRE发育中的作用以及CREB对DRE中GABA相关受体的影响。
方法：慢病毒转染使正常大鼠海马CREB的表达升高或降低，然后接受了锂-毛果芸香碱诱导的癫痫模型。苯巴比妥(PB)钠和卡马西平(CBZ)用于选择耐药癫痫模型。采用westernblot和荧光定量PCR检测大鼠海马GABAA受体α1、β2和γ2亚基及CREB蛋白的表达水平。
结果：与对照组相比，过表达组的癫痫发作频率和持续时间增加。此外,严重程度,频率,在表达减少的组中，癫痫发作的持续时间减少。CREB蛋白和CREBmRNA表达水平的海马分析产生了类似的发现。改变大鼠海马中的CREB蛋白表达可以负向调节GABAA受体α1，β2和γ2的表达和转录水平，这表明CREB可以作为开发癫痫治疗方案和药物的潜在靶标。
结论：我们的研究表明，增强的CREB表达促进DRE的发展，并负向调节GABAA受体水平，并且抑制CREB表达可能会降低DRE的发生率。
目的：本研究探讨了CREB在DRE发育中的作用以及CREB对DRE中GABA相关受体的影响。
方法：慢病毒转染使正常大鼠海马CREB的表达升高或降低，然后接受了锂-毛果芸香碱诱导的癫痫模型。苯巴比妥(PB)钠和卡马西平(CBZ)用于选择耐药癫痫模型。采用westernblot和荧光定量PCR检测大鼠海马GABAA受体α1、β2和γ2亚基及CREB蛋白的表达水平。
结果：与对照组相比，过表达组的癫痫发作频率和持续时间增加。此外,严重程度,频率,在表达减少的组中，癫痫发作的持续时间减少。CREB蛋白和CREBmRNA表达水平的海马分析产生了类似的发现。改变大鼠海马中的CREB蛋白表达可以负向调节GABAA受体α1，β2和γ2的表达和转录水平，这表明CREB可以作为开发癫痫治疗方案和药物的潜在靶标。
结论：我们的研究表明，增强的CREB表达促进DRE的发展，并负向调节GABAA受体水平，并且抑制CREB表达可能会降低DRE的发生率。
