安神定志灵对ADHD模型大鼠前额叶、纹状体CDK5/DARPP32/PP1信号通路的影响

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(P0.05,P0.01),PP1和DARPP32的表达水平明显升高(P0.05,P0.01)。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(P0.05,P0.01),PP1和DARPP32的表达水平明显降低(P0.05,P0.01)。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。

Effects of Anshen Dingzhiling Compound on Signaling Pathway of CDK5/DARPP32/PP1 in ADHD Rats' Striatum and Prefrontal Cortex

Objective: To investigate the effects of Anshen Dingzhiling (ADL) compound on signaling pathways of cyclin-dependent kinase 5(CDK5)/dopamine and adenosine 3'', 5''-monophosphate-regulated phospho-protein, 32 kDa (DARPP-32)/protein phosphatase 1(PP1) in striatum and prefrontal cortex of spontaneously hypertensive rats (SHR) with attention deficit hyperactivity disorder (ADHD). Method: The 10 Wistar Kyoto rats were used as normal group. 50 SHR rats were randomly divided into model group, methylphenidate (MPH) group (2 mg · kg^-1by gavage), and ADL low, middle, and high dose groups (6.7, 13.4, 26.7 g · kg^-1 respectively), n=10 in each group. The rats in various groups received corresponding drugs by ig administration, bid. The brain tissues of the rats were taken after 4 weeks of treatment. Real-time PCR, Western blot and immunohistochemistry were used to detect the protein and mRNA expression levels of CDK5, p35, DARPP32, PP1 and cAMP response element binding protein (CREB) in striatum and prefrontal cortex of SHR rats. Result: As compared with normal group, the protein and mRNA expression levels of CDK5, p35 and CREB in striatum and prefrontal cortex were significantly reduced in rats of model group (P<0.05, P<0.01), the expression levels of DARPP32 and PP1 were significantly increased (P<0.05, P<0.01). After treatment, the protein and mRNA expression levels of CDK5, p35 and CREB in striatum and prefrontal cortex were significantly increased in rats of MPG group and ADL groups as compared with the model group (P<0.05, P<0.01), while the expression levels of DARPP32 and PP1 were significantly reduced (P<0.05, P<0.01). Conclusion: The acts of SHR rats may be associated with the signaling pathway of CDK5/DARPP32/PP1.ADL can exert the therapeutic action by regulating the signaling pathway of CDK5/DARPP32/PP1.