柴胡疏肝散通过mir124调控海马神经元内p-CREB、BDNF表达的机制研究

Study on the Mechanism of Chaihu Shugan Powder Mediated mir124 Regulation of the Expression of BDNF and p-CREB Protein in Hippocampal Neurons

目的:探索Mir124对大鼠海马神经元内p-CREB、BDNF蛋白的影响和作用机制以及中药柴胡疏肝散对此通路的干预作用。方法:培养海马神经元,并予中药柴胡疏肝散含药血清干预,转染rno-mir124建立mir124高表达系,实时定量RT-PCR检测mir124表达水平,Western-blot检测p-CREB、BDNF蛋白表达。结果:海马神经元经转染rno-mir124后成功建立了mir124高表达系,转染后p-CREB和BDNF蛋白表达降低,差异有显著性;而予柴胡疏肝散含药血清处理可抑制mir124表达,并进一步降低p-CREB,但空白血清加转染mir124组与柴胡疏肝散处理加转染mir124组之间比较,BDNF蛋白表达差异无统计学意义。结论:海马神经元内mir124表达升高,可以降低CREB磷酸化,进一步降低BDNF蛋白表达,从而影响海马神经元的可塑性,而中药柴胡疏肝散可以抑制mir124的表达,促进CREB磷酸化。

Objective： To explore the influence of the mir124 on p-CREB and BDNF protein in hippocampal neurons, and the intervention effect of Chaihu Shugan Powder on this pathyway. Methods ：＇ Cultured hippoeampal neurons, and treated by ser- um containing Chaihu Shugan Powder, transfection of rno-mir124 to establish a high expression system of mir124; detection of mir124 expression level with real-time quantitative RT-PCR, and analysis of CREB, BDNF protein expression by Western blot. Results ： miH24 high expression system was successfully established in the hippocampal neurons by transfection of rno-mir124. In the group of transfection rno-mir124, the expression of p-CREB and BDNF was lower than that of the controlled group, while treated by serum containing Chaihu Shugan Powdwer, the expression of p-CREB protein was significantly higher than that of the group which were transfection of rno-mir124. Conclusion： The high expression of mir124 in hippocampusneurons can inhibite CREB phospborylafion and further reduce the expression of BDNF protein, which may affect the plasticity of hippocampal neu- rons, and Chaihu Shugan Powder may be regulate this pathway.