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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 effect and mechanism of Mir124 on p-CREB and BDNF protein in rat hippocampal neurons and the effect of Chaihushugan decoction on this pathway. Methods: The hippocampal neurons were cultured and treated with the Chinese medicine Chaihu Shugan San serum. The mir124 overexpression lines were transfected with rno-mir124. The expression of mir124 was detected by real-time quantitative RT-PCR. The expression of p-CREB and BDNF Protein. Results: The mir124 overexpression line was successfully established in hippocampal neurons after transfected with rno-mir124, the expression of p-CREB and BDNF protein decreased after transfection, and the difference was significant; however, the treatment with Chaihu Shugan San serum could inhibit mir124 and further reduce the expression of p-CREB. However, there was no significant difference in the expression of BDNF between blank serum plus transfected mir124 group and Chaihu Shugan decoction plus mir124 group. CONCLUSION: Increased mir124 expression in hippocampal neurons can decrease CREB phosphorylation and further decrease BDNF protein expression, thereby affecting hippocampal neuronal plasticity. However, Chaihu Shugan San inhibits the expression of mir124 and promotes CREB phosphorylation.