帕罗西汀通过Akt/GSK3β/CREB通路对焦虑大鼠行为学的改善作用

Paroxetine in improving behaviors of anxious rats through Akt/GSK3β/CREB pathway

目的基于蛋白激酶B(Akt)/糖原合成酶激酶3β(GSK3β)/环腺苷酸反应元件结合蛋白(CREB)通路探讨帕罗西汀对焦虑大鼠行为学的影响。方法焦虑大鼠模型采用单笼饲养且接受不确定性空瓶饮水刺激方法建立。SPF级SD雄性大鼠随机分为对照组、模型组、低中高剂量帕罗西汀组(帕罗西汀3 mg/kg、6 mg/kg、12 mg/kg)、Akt抑制剂组(Akt抑制剂Perifosine 20 mg/kg+帕罗西汀12 mg/kg),每组6只。观察大鼠攻击、探究、修饰行为;高架十字迷宫(EPM)试验记录大鼠进入开放臂次数(OE)、进入开放臂时间(OT)、进入封闭臂次数(CE)、进入封闭臂时间(CT)、向下探究次数(HD),并计算OE+CE、OE比例(OE%)、OT比例(OT%);实时荧光定量PCR法(qRT-PCR)和免疫印迹法(Western blot)检测大鼠海马组织中Akt、GSK3β、CREB mRNA及蛋白水平。结果与对照组比较,模型组大鼠毛色干枯无光、粗糙,易被激怒,探究行为、修饰行为、攻击行为水平升高(P<0.05),OE+CE、OE%、OT%、HD水平降低(P<0.05),p-Akt/Akt、p-GSK3β/GSK3β、p-CREB/CREB蛋白水平均降低(P<0.05)。与模型组比较,低中高剂量帕罗西汀组、Akt抑制剂组大鼠给药后状态均有不同程度改善,毛色逐渐光泽,活动变多,探究行为、修饰行为、攻击行为水平降低(P<0.05),OE+CE、OE%、OT%、HD水平升高(P<0.05),p-Akt/Akt、p-GSK3β/GSK3β、p-CREB/CREB蛋白水平均升高(P<0.05)。且帕罗西汀处理基础上使用Akt抑制剂可部分逆转帕罗西汀对Akt/GSK3β/CREB通路的激活作用及对焦虑样行为的改善作用。结论帕罗西汀可通过激活海马组织中Akt/GSK3β/CREB通路,改善大鼠焦虑样行为。

Objective To explore effects of paroxetine in improving behaviors of anxious rats based on protein kinase B(Akt)/glycogen synthase kinase 3β(GSK3β)/cyclic adenosine monophosphate(cAMP)response-element binding protein(CREB)pathway.Methods Anxious rat model for rats which were fed with single cages and received an indeterminate empty bottle drinking water stimulus method was established.SPF SD male rats were randomized to the control group,model group,and low-medium-,high-dose paroxetine group(paroxetine in 3mg/kg,6mg/kg,12mg/kg),Akt inhibitor group(Perifosine in 20mg/kg+paroxetine in 12mg/kg)with 6 rats in each group.Behaviors including the aggression,exploration and modification were observed.Open arm entries(OE),open arm time(OT),close arm entry(CE)close arm entry time(CT)and head-dipping(HD)were recorded in the elevated plus maze(EPM)test,and OE+CE,OE%and OT%were calculated.The mRNA and protein level of Akt,GSK3βand CREB in hippocampus were detected by real-time fluorescence quantitative PCR(qRT-PCR)and Western blot(WB)Results Compared to the control group,the hair color of rats in the model group was dry,dull,rough.The rats were vulnerable to be irritated,and the behavior levels of exploratory,modification and aggressive increased significantly(P<0.05).The level of OE+CE,OE%,OT%and HD decreased significantly(P<0.05).The protein level of p-Akt/Akt,p-GSK3β/GSK3βand p-CREB/CREB decreased significantly(P<0.05).Compared to the model group,the improvement conditions in Paroxetine groups and Akt inhibitor group were found after administration.The hair color of rats was gradually glossy,and the activities increased.The behavior level ofexploratory,modification and aggressive decreased significantly(P<0.05).The level of OE+CE,OE%,OT%and HD increased significantly(P<0.05).The protein level of p-Akt/Akt,p GSK3β/GSK3βand p CREB/CREB increased significantly(P<0.05).In addition,on the basis of paroxetine treatment,Akt inhibitor could partially reverse the activation effect of paroxetine on Akt/GSK3β/CREB pathway and improvement effect on anxiety-like behavior.Conclusion Paroxetine can improve the anxiety-like behavior by activating the Akt/GSK3β/CREB pathway in the hippocampus.