中脑腹侧被盖区SIRT1过表达对海洛因依赖小鼠学习记忆和突触结构可塑性的影响  

Effect of SIRT1 overexpression in ventral tegmental area on learning,memory and synaptic structural plasticity in heroin-dependent mice

作　　者：马旋 谭雨维 刘美君 郑欣雅 赵传睿 尹丹 王茜 李豪 夏白娟 MA Xuan;TAN Yuwei;LIU Meijun;ZHENG Xinya;ZHAO Chuanrui;YIN Dan;WANG Qian;LI Hao;XIA Baijuan(Department of Histology and Embryology,School of Basic Medical Sciences,Guizhou Medical University,Guian 561113,Guizhou,China;Department of Outpatient,Guizhou Provincial General Hospital of the Chinese Armed Police Force,Guiyang 550004,Guizhou,China;Key Laboratory for Research on Autoimmune Diseases of Higher Education Schools in Guizhou Province,Guian 561113,Guizhou,China)

机构地区：[1]贵州医科大学基础医学院组织学与胚胎学教研室,贵州贵安561113 [2]武警贵州省总队医院门诊部,贵州贵阳550004 [3]贵州省普通高校自身免疫病研究重点实验室,贵州贵安561113

出　　处：《贵州医科大学学报》2024年第11期1601-1608,共8页Journal of Guizhou Medical University

基　　金：国家自然科学基金(81960253);贵州省高层次留学人才项目(2020-02);贵州省卫生健康委科学技术基金项目(gzwkj2021-511);贵州医科大学国基培育项目(20NSP014);2020年国家大学生创新创业计划训练项目(202010660031);贵州省中医药管理局中医药、民族医药科学技术研究课题(QZYY-2021-039)

摘　　要：目的探讨中脑腹侧被盖区(VTA)过表达沉默信息调节因子1(SIRT1)对海洛因依赖小鼠学习记忆能力和突触结构可塑性的影响。方法42只C57BL/6J小鼠分为生理盐水对照组(SCG组,n=14)、海洛因依赖组(HDG组,n=14)以及SIRT1过表达海洛因依赖组(SIRT1-OE+H组,n=14),后两组采用逐日递增注射海洛因液体建立海洛因依赖小鼠模型,SIRT1-OE+H组需先VTA注射rAAV9-rSIRT1腺相关病毒;观察小鼠条件位置偏爱(CPP)及Morris水迷宫(MWM)学习记忆行为;转染rAAV9-rSIRT121 d时,透射电镜观察VTA脑区突触超微结构,蛋白质印迹(Western blot)检测小鼠VTA脑区突触可塑性相关蛋白的表达。结果CPP试验中,与SCG相比较,HDG组小鼠在伴药箱中停留延长、CPP评分增高,SIRT1-OE+H组改变更加显著(P<0.05);MWM的定位航行试验结果显示,第4天和第5天,SIRT1-OE+H组逃避潜伏期延长(P<0.05);空间探索试验结果显示,HDG与SIRT1-OE+H组小鼠穿越平台次数增多(P<0.05),但目标象限停留时间比率、目标象限路程比率差异无统计学意义(P>0.05);透射电镜结果显示,HDG和SIRT1-OE+H组突触后膜致密物增厚,突触界面曲率增大;Western blot结果显示SIRT1-OE+H组中突触素(Syn)、突触后致密区蛋白95(PSD95)、生长相关蛋白43(Gap43)表达较SCG明显增多(P<0.05)。结论SIRT1参与了海洛因依赖过程,其机制可能是SIRT1过表达增强了突触可塑性,促进了海洛因引起的奖赏效应,但SIRT1对空间学习记忆能力的影响不显著。Objective To investigate the effect of silent mating type information regulation 1(SIRT1)overexpression in the ventral tegmental area(VTA)of the midbrain on learning,memory capacities and synaptic structural plasticity in heroin-dependent mice.Methods A total of 42 C57BL/6J mice were evenly divided into three groups:saline control group(SCG,n=14),heroin-dependent group(HDG,n=14)and SIRT1 overexpression in heroin-dependent group(SIRT1-OE+H,n=14).The latter two groups were established for a heroin-dependent mouse model by daily increasing injection of heroin solution.SIRT1-OE+H group were given with rAAV9-rSIRT1 adenovirus by VTA injection first.Learning and memory behaviors were monitored using conditioned place preference(CPP)and Morris water maze(MWM).On 21 st day after rAAV9-rSIRT121 d infection,the ultrastructure of VTA synapses was observed by transmission electron microscopy.Western blot was performed to detect the expression of synaptic plasticity-related proteins in VTA.Results CCP showed that HDG group stayed longer in the drug box and had higher CPP scores than those in SCG,while SIRT1-OE+H group showed more significant changes when compared to SCG(P<0.05).MWM localization navigation test revealed that the escape latency was prolonged in SIRT1-OE+H group on the 4th and 5th days.Spatial exploration test showed that the numbers of the crossing the platform in HDG and SIRT1-OE+H groups were increased(P<0.05),but there were no statistical differences in the ratio of the target quadrant run to the total run and the ratio of target quadrant time to total time(P>0.05).Transmission electron microscopy results showed that thicken postsynaptic membrane density and increased curvature of the synaptic interface in HDG and SIRT1-OE+H groups.Western blot results showed that Syn,PSD95,and Gap43 expressions were significantly enhanced in SIRT1-OE+H group when compared to SCG(P<0.05).Conclusion SIRT1 mediates the process of heroin dependence during the time course of dependence.The mechanism may be that overexpression of

关 键 词：沉默信息调节因子1 中脑腹侧被盖区 记忆 突触可塑性 海洛因 

分 类 号：R329.3[医药卫生—人体解剖和组织胚胎学]