中央杏仁核GABA神经元对小鼠情绪和摄食的调控研究  

GABA neurons in central amygdala regulate emotional behavior and food intake in mice

作　　者：刘宁[1,2] 纪鹏飞[3] 胡阳 于立政 张鸿飞 郭菲菲 LIU Ning;JI Pengfei;HU Yang;YU Lizheng;ZHANG Hongfei;GUO Feifei(Department of Physiology and Pathophysiology,School of Basic Medicine,Qingdao University,Qingdao 266071,Chi-na;Department of Nursing,Heze Medical College,Heze 274000,China;Qingdao Medical College,Qingdao University,Qingdao 266071,China)

机构地区：[1]青岛大学基础医学院生理学与病理生理学系,山东青岛266071 [2]菏泽医学专科学校护理系,山东菏泽274000 [3]青岛大学青岛医学院,山东青岛266071

出　　处：《中国病理生理杂志》2023年第4期588-598,共11页Chinese Journal of Pathophysiology

基　　金：中国博士后科研基金资助项目(No.2018M632627);国家级大学生创新创业训练计划项目(No.202111065268)。

摘　　要：目的:探讨中央杏仁核(CeA)内γ-氨基丁酸(GABA)能神经元到下丘脑室旁核(PVN)神经通路对小鼠情绪和摄食的调控作用及潜在机制。方法:采用慢性束缚应激(CRS)建立KM小鼠焦虑抑郁样模型。(1)在CRS小鼠和正常对照(NOR)小鼠的PVN注射0.3µL生理盐水(NS)或GABAA受体拮抗剂荷包牡丹碱(BIC),并据此将小鼠分为4组(n=10):NOR+NS组、NOR+BIC组、CRS+NS组和CRS+BIC组。通过旷场实验、高架十字迷宫实验、大理石掩埋实验和强迫游泳实验观察小鼠行为学的变化;观察2周内小鼠摄食和体重的变化。(2)采用荧光金逆行追踪结合荧光免疫组织化学法检测CeA-PVN神经通路。(3)采用化学遗传法将腺相关病毒载体AAV2/9-mDlx-hM3D(Gq)-EGFP-ER2-WPRE-PA定向注射于CeA,小鼠分为4组(n=10):NS(腹腔注射)+NS(PVN微量注射)组、NS+BIC组、氯氮平N-氧化物(CNO)+NS组和CNO+BIC组。观察CeA-PVN的GABA能神经通路对小鼠行为学和摄食体重的影响。结果:行为学结果显示,CRS+NS组出现焦虑抑郁样行为改变;与NOR+NS组相比,CRS+NS组小鼠在6~12 h、12~24 h和2周内的摄食和体重显著降低(P<0.05)。PVN注射BIC后CRS小鼠焦虑抑郁样行为显著减轻(P<0.05),6~12 h、12~24 h和2周内的摄食量显著增加(P<0.05),持续给药第8天开始CRS小鼠的体重显著增加(P<0.05)。荧光金逆行追踪结合免疫荧光组织化学实验结果显示,CeA的GABA神经元可以投射至PVN。化学遗传法和行为学实验结果显示,与NS+NS组相比,CNO+NS组小鼠出现焦虑抑郁样行为(P<0.05),在6~12 h、12~24 h和2周内的摄食量显著降低(P<0.05),且第6天开始体重显著减轻(P<0.05);与CNO+NS组相比,CNO+BIC组的焦虑抑郁样行为显著减轻(P<0.05),在6~12 h、12~24 h和2周内的摄食量显著增加(P<0.05),给药第8天体重也开始显著增加(P<0.05)。结论:CeA-PVN的GABA能神经通路可参与小鼠的情绪行为和摄食调控。AIM:To investigate the effects ofγ-aminobutyric acid(GABA)-ergic neural pathway from the central amygdala(CeA)to the paraventricular nucleus(PVN)on the regulation of emotional behavior and food intake in mice and its potential mechanisms.METHODS:Chronic restraint stress(CRS)was used to establish a KM mouse model of anxiety and depression.(1)The mice were randomly divided into 4 groups(n=10):normal(NOR)mice+normal saline(NS)group,NOR+GABAA receptor antagonist bicuculline(BIC)group,CRS+NS group and CRS+BIC group.A total of 0.3µL of NS or BIC was injected into the PVN of CRS mice or NOR mice through a pre-buried catheter.Open-field test,elevated plus maze test,marble burying test and forced swimming test were used to observe the behavior of anxiety and depression model mice.The changes of food intake and body weight within 2 weeks after the injection were monitored.(2)·588·Fluorogold(FG)retrograde tracing combined with fluorescence immunohistochemistry was used to detect the CeA-PVN neural pathways.(3)The mice pre-injected with AAV2/9-mDlx-hM3D(Gq)-EGFP-ER2-WPRE-PA in the CeA were randomly divided into 4 groups(n=10):NS(intraperitoneal injection)+NS(PVN microinjection)group,NS+BIC group,clozapine N-oxide(CNO)+NS group,and CNO+BIC group.The effects of GABAergic neural pathways from CeA to PVN on emotional behavior,food intake and body weight were detected.RESULTS:The results of behavioral tests showed that the mice in CRS+NS group had anxiety-and depression-like behavior.Compared with NOR+NS group,the food intake and body weight decreased significantly within 6~12 h,12~24 h and 2 weeks in CRS+NS group(P<0.05).After the microinjection of BIC into the PVN,the anxiety-and depression-like behavior of CRS mice were alleviated(P<0.05),the food intake increased significantly at 6~12 h,12~24 h and 2 weeks(P<0.05),and the body weight of CRS mice increased considerably from the 8th day of continuous administration(P<0.05).Results of FG retrograde tracing combined with fluorescence immunohistochemistry showed that GABA neur

关 键 词：中央杏仁核 下丘脑室旁核 Γ-氨基丁酸 情绪 摄食 

分 类 号：R749.72[医药卫生—神经病学与精神病学] R363.2[医药卫生—临床医学]