下丘脑外侧区-伏隔核黑色素浓缩激素神经通路对小鼠焦虑样行为的调控作用  

The regulatory role of the hypothalamic lateral area-ventral tegmental nucleus melanin-concentrating hormone neural pathway in anxiety-like behavior in mice

作　　者：李颜君 薛佳淼[1] 桑宇轩 周艳 郜慧英[2] 张娜娜[3] Li Yanjun;Xue Jiamiao;Sang Yuxuan;Zhou Yan;Gao Huiying;Zhang Nana(Qingdao Medical College,Qingdao University,Qingdao 266071,China;Department of Nursing,Heze Medical College,Heze 274000,China;Department of Clinical Laboratory,the Afiliated Hospital of Qingdao University,Qingdao 266003,China)

机构地区：[1]青岛大学青岛医学院,青岛266071 [2]菏泽医学专科学校护理系,菏泽274000 [3]青岛大学附属医院检验科,青岛266003

出　　处：《中华行为医学与脑科学杂志》2024年第10期876-882,共7页Chinese Journal of Behavioral Medicine and Brain Science

基　　金：青岛市医药卫生科研指导项目(2022-WJZD108,2023-WJZD106)。

摘　　要：目的探讨下丘脑外侧区(lateral hypothalamus,LHA)至伏隔核(nucleus accumbens,NAc)间黑色素浓缩激素(melanin-concentrating hormone,MCH)神经通路对小鼠焦虑样行为的调控作用。方法37只6~8周龄SPF级雄性C57BL/6J小鼠(18~22 g)进行以下实验:(1)5只小鼠,NAc内注射荧光金(flouro-gold,FG),1周后采用逆行追踪结合免疫荧光染色法,观察LHA中FG和MCH免疫阳性神经元共存情况。(2)32只小鼠LHA注射可激活MCH神经元的腺相关病毒,2周后按体质量匹配原则分为4组:NS+NS组[腹腔注射生理盐水(normal saline,NS),NAc注射NS(1.5μL)]、NS+SNAP94847组[腹腔注射NS,NAc注射SNAP(2 mg/mL,1.5μL),SNAP94847为MCH 1型受体拮抗剂]、CNO+NS组[腹腔注射氯氮平N-氧化物(CNO,0.15 mg/kg),NAc注射NS(1.5μL)]和CNO+SNAP组[腹腔注射CNO,NAc注射SNAP]。采用旷场实验(open field test,OFT)、高架十字迷宫(elevated plus maze,EPM)实验和埋珠实验(marble burial test,MBT)观察化学遗传学方法激活MCH神经元对小鼠焦虑行为的影响。采用Graphpad Prism 8.0版软件进行统计分析,多组间比较采用2×2析因设计方差分析。结果(1)FG逆行追踪结合荧光免疫组化方法结果显示,LHA内MCH神经元发出神经纤维投射至NAc神经元。(2)化学遗传法激活MCH神经元后,各组小鼠OFT实验中,CNO干预和SNAP干预对中心区域停留时间(F_(交互)=2.899,P>0.05)和移动距离(F_(交互)=1.603,P>0.05)的交互作用均无显著性。中心区域停留时间(F_(CNO)=6.767,F_(SNAP)=7.656,均P<0.05)和移动距离(F_(CNO)=12.480,F_(SNAP)=7.999,均P<0.01)的CNO干预主效应和SNAP干预主效应均显著。CNO+NS组小鼠中心区域停留时间[(89.00±19.16)s,(63.75±13.58)s,P<0.05]和移动距离[(593.79±108.18)cm,(426.81±66.14)cm,P<0.05]显著短于NS+NS组(均P<0.05);CNO+SNAP组中心区域停留时间[(87.38±16.57)s]和移动距离[(569.27±73.20)cm]显著长于CNO+NS组(均P<0.05)。EPM实验中,CNO干预和SNAP干预对小鼠进入开放臂次数(F_(交互)=2.79)和�Objective:To explore the regulatory role of the melanin-concentrating hormone(MCH)neural pathway from the lateral hypothalamus(LHA)to the nucleus accumbens(NAc)in modulating anxiety-like behavior in mice.Methods:Totally 37 male SPF-grade C57BL/6J mice,aged 6 to 8 weeks and weighed 18-22 g,were used for the following experiment:(1)Five mice received an injection of Fluoro-Gold(FG)into the NAc,followed by retrograde tracing combined with immunofluorescence staining after one week to observe the coexistence of FG and MCH immunopositive neurons in the LHA.(2)Thirty-two mice were injected with adeno-associated virus which could activate the MCH neurons into the LHA.After two weeks,they were randomly divided into four groups and received different drug injections:normal saline(NS,intraperitoneal injection)+NS(1.5μL,injection in the NAc)group,NS(intraperitoned injection)+SNAP94847(SNAP,2 mg/mL,1.5μL,injection in the NAc)group,chlorprothixene N-oxide(CNO,0.15 mg/kg,intraperitoned injection)+NS(1.5μL,injection in the NAc)group,and CNO(intraperitoned injection)+SNAP(1.5μL injection in the NAc)group,and the SNAP94847 was an antagonist for MCH typel receptor.The open field test(OFT),elevated plus maze(EPM),and marble burial test(MBT)were employed to assess the impact of chemogenetic activation of MCH neurons on anxiety-related behavior in mice.Results:(1)The results of FG retrograde tracing combined with immunofluorescence histochemistry showed that MCH neurons in the LHA project their neural fibers to NAc neurons.(2)In the chemogenetic experiment,there was no significant interaction effect between CNO and SNAP in terms of the duration of stay(F_(interaction)=2.899,P>0.05)and the distance moved(F_(interaction)=1.603,P>0.05)in the central area during OFT experiments.However,the main effects of both CNO and SNAP intervention were significant in the duration of stay(F_(CNO)=6.767,F_(SNAP)=7.656,both P<0.05)and the distance moved(F_(CNO)=12.480,F_(SNAP)=7.999,both P<0.01)in the central area.Compared to the NS+NS group,mice i

关 键 词：下丘脑外侧区 伏隔核 黑色素浓缩激素 黑色素浓缩激素受体拮抗剂 SNAP94847 焦虑 小鼠 

分 类 号：R749.72[医药卫生—神经病学与精神病学]