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目的:探讨中药复方安神定志灵对注意缺陷多动障碍(ADHD)模型自发性高血压大鼠(SHR)前额叶、纹状体中细胞周期素依赖性蛋白激酶5(CDK5)/32k Da多巴胺与环磷酸腺苷(c AMP)调节的磷蛋白(DARPP32)/蛋白磷酸酶1(PP1)信号通路的影响。方法:将50只SHR大鼠运用随机区组设计分为模型组、利他林组(2 mg·kg-1)、安神定志灵低、中、高剂量(6.7,13.4,26.7 g·kg-1)组,每组10只,另设10只Wistar京都大鼠为正常组。各组ig相应药物,每日2次,治疗4周后取大鼠脑组织。采用蛋白质免疫印迹法(Western blot),免疫组织化学法及实时荧光定量聚合酶链式反应(Real-time PCR)检测各组大鼠前额叶和纹状体中CDK5,调节蛋白35(p35),DARPP32,PP1和c AMP反应元件结合蛋白(CREB)蛋白和mRNA表达水平。结果:与正常组比较,模型组大鼠前额叶、纹状体中CDK5,p35及CREB的蛋白和mRNA表达水平均明显降低(P<0.05,P<0.01),PP1和DARPP32的表达水平明显升高(P<0.05,P<0.01)。经治疗后,与模型组比较,利他林组、安神定志灵各剂量组前额叶、纹状体中CDK5,p35,CREB蛋白和mRNA表达水平明显升高(P<0.05,P<0.01),PP1和DARPP32的表达水平明显降低(P<0.05,P<0.01)。结论:SHR大鼠行为表现与CDK5/DARPP32/PP1信号通路密切相关,安神定志灵可以通过调控该信号通路中相关因子的表达发挥治疗作用。
OBJECTIVE: To investigate the effects of Anshen Dingzhi Ling on the expression of cyclin-dependent kinase 5 (CDK5) / 32kDa dopamine in the prefrontal lobe and striatum of spontaneous hypertensive rats (SHR) model of attention deficit hyperactivity disorder (ADHD) And cAMP regulated phosphoprotein (DARPP32) / protein phosphatase 1 (PP1) signaling pathways. Methods: Fifty SHR rats were randomly divided into model group, Ritalin group (2 mg · kg -1), Anshen Dingzhi Ling low, middle and high doses (6.7, 13.4 and 26.7 g · kg- 1) group, with 10 rats in each group. Another 10 Wistar Kyoto rats were normal rats. Each group of ig corresponding drugs, 2 times a day, 4 weeks after treatment, rat brain tissue. Western blot, immunohistochemistry and real-time PCR were used to detect the expression of CDK5, regulatory protein 35 (p35) in prefrontal and striatum, DARPP32, PP1 and c AMP response element binding protein (CREB) protein and mRNA expression levels. Results: Compared with normal group, the protein and mRNA expressions of CDK5, p35 and CREB in prefrontal and striatum of model group were significantly decreased (P <0.05, P <0.01), and the expression levels of PP1 and DARPP32 were significantly increased High (P <0.05, P <0.01). After treatment, compared with the model group, the expressions of CDK5, p35, CREB protein and mRNA in the frontal lobe and striatum were significantly increased (P <0.05, P <0.01) PP1 and DARPP32 expression levels were significantly lower (P <0.05, P <0.01). Conclusion: The behavior of SHR rats is closely related to the CDK5 / DARPP32 / PP1 signaling pathway. Anshen Dingzhi Ling can play a therapeutic role by regulating the expression of related factors in this signaling pathway.