海马AKT/GSK3β/CRMP2信号通路改变对大鼠抑郁样行为及微管蛋白的影响  

Effects of hippocampal AKT/GSK3β/CRMP2 signaling pathway on depression-like behavior and tubulin in rats

作　　者：魏艳艳 陈景旭[1] 肖玲[2] 吴作天[2] 何静 张楠 王高华 Wei Yanyan;Chen Jingxu;Xiao Ling;Wu Zuotian;He Jing;Zhang Nan;Wang Gaohua(Department of Affective Disorder,Beijing Hui-Long-Guan Hospital,Peking University,Beijing 100096,China;Renmin Hospital of Wuhan University,Wuhan 430060,China)

机构地区：[1]北京回龙观医院情感障碍科,100096 [2]武汉大学人民医院精神卫生中心,430060

出　　处：《中华行为医学与脑科学杂志》2020年第8期713-718,共6页Chinese Journal of Behavioral Medicine and Brain Science

基　　金：国家自然科学基金资助项目(81571325;81871072);国家科技支撑计划(2015BAI13B01)。

摘　　要：目的研究磷脂酰肌醇3激酶(phosphatidylinositol 3 kinase,PI3K)抑制剂LY294002对AKT/GSK3β/CRMP2信号通路的影响,及通路改变对大鼠抑郁样行为及微管蛋白的影响。方法16只成年雄性SD大鼠采用盲法随机分为LY294002组及溶剂DMSO组,每组8只。麻醉后进行海马CA1区脑立体置管,置管1周后给予LY294002或DMSO脑立体注射,并进行抑郁样行为检测。采用RT-qPCR技术检测海马AKT、GSK3β、CRMP2、tubulin的mRNA表达水平,采用Western blot检测海马AKT、p-AKT、GSK3β、p-GSK3β、CRMP2、p-CRMP2以及微管动态性相关蛋白Acet-tubulin、Tyr-tubulin表达水平。比较两组大鼠抑郁样行为表现及分子表达差异。结果旷场实验显示,LY294002组大鼠的中心移动距离较DMSO组显著下降[(3.64±2.17)cm;(31.51±12.68)cm;t=2.69,P=0.03];LY294002组中心区持续时间较DMSO组亦显著下降[(0.73±0.46)s;(4.85±2.10)s;t=2.33,P=0.04]。强迫游泳测试结果显示LY294002组大鼠的不动时间有升高的趋势,但差异无统计学意义。糖水偏好实验显示两组大鼠糖水消耗量差异无统计学意义(P>0.05)。RT-qPCR结果显示,LY294002组大鼠海马组织CRMP2 mRNA表达水平显著下降(P<0.05)。Western blot结果显示,与DSMO组比较,LY294002组大鼠海马组织p-AKT及p-GSK3β表达水平下降(P<0.05);CRMP2表达水平降低,p-CRMP2表达水平显著升高,同时,Tyr-tubulin表达水平下降,Acet-tubulin表达水平显著升高,两组均差异有统计学意义(均P<0.05)。结论LY294002通过影响AKT/GSK3β/CRMP2信号通路,导致微管动态性损害,并使大鼠出现抑郁样行为。Objective To study the effects of LY294002,a phosphatidylinositol 3 kinase(PI3K)inhibitor,on the AKT/GSK3β/CRMP2 signaling pathway,and the effect of pathway alterations on the depression-like behavior and microtubulin proteins of SD rats.Methods Sixteen adult male SD rats were randomly divided into LY294002 group and solvent DMSO group by blind method,8 rats in each group.After anesthesia,the rats were subjected to stereotactic tube placement in the hippocampal CA1 area.After 1 week of the tube placement,stereoscopic injection of LY294002 or DMSO was given,and depression-like behaviors were detected.RT-qPCR technology was used to detect hippocampal AKT,GSK3β,CRMP2,and tubulin mRNA expression levels,and Western blot technology was used to detect the expression levels of hippocampal AKT,p-AKT,GSK3β,p-GSK3β,CRMP2,p-CRMP2,and microtubule dynamic-related proteins markers Acet-tubulin,Tyr-tubulin.Analysis and comparison of behavioral and molecular expression differences between the two groups were conducted.Results The open field experiment showed that the central movement distance of the rats in the LY294002 group were significantly lower than those in the control group((3.64±2.17)cm,(31.51±12.68)cm;t=2.69,P=0.03),and the duration of the central area in the LY294002 group were also significantly lower than those in the control group((0.73±0.46)s,(4.85±2.10)s;t=2.33,P=0.04).The results of the forced swimming test showed that the immobility time of rats in the LY294002 group had upward trend,but the difference was not statistically significant.The sucrose preference results showed no significant difference in sugar water consumption between the two groups(P>0.05).The RT-qPCR results showed that the expression level of CRMP2 mRNA in the LY294002 group decreased significantly(P<0.05).Western blot results showed that compared with solvent DMSO group,the expression levels of p-AKT and p-GSK3βin the LY294002 group decreased(P<0.05),the expression level of CRMP2 was decreased and the expression level of p-CRMP2 was s

关 键 词：PI3K抑制剂 脑衰反应调节蛋白2 信号通路 微管蛋白 抑郁 

分 类 号：R749.4[医药卫生—神经病学与精神病学]