星状神经节阻滞对创伤后应激障碍模型小鼠条件性恐惧记忆的影响及其神经环路机制  

Effect and neural circuit mechanism of stellate ganglion block on conditioned fear memory in post-traumatic stress disorder model mice

作　　者：王子恒 刘洲亮 王婷婷 朱杨子 王立伟 Wang Ziheng;Liu Zhouliang;Wang Tingting;Zhu Yangzi;Wang Liwei(Jiangsu Province Key Laboratory of Anesthesiology,Xuzhou Medical University,Xuzhou 221004,China;Xuzhou Clinical School,Xuzhou Medical University,Xuzhou 221004,China)

机构地区：[1]徐州医科大学江苏省麻醉学重点实验室,徐州221004 [2]徐州医科大学徐州临床学院,徐州221004

出　　处：《中华行为医学与脑科学杂志》2024年第4期295-302,共8页Chinese Journal of Behavioral Medicine and Brain Science

基　　金：国家自然科学基金(81801332);江苏省老年健康科研基金(LKZ2023016)。

摘　　要：目的:探讨星状神经节阻滞(stellate ganglion block,SGB)对创伤后应激障碍(post-traumatic stress disorder,PTSD)模型小鼠条件性恐惧记忆的调控作用及其机制。方法:选取8~9周龄SPF级雄性C57BL/6J小鼠,根据随机数字表法分组,并完成以下实验:(1)32只小鼠分为PTSD组、PTSD+七氟烷组(Sev组)、PTSD+生理盐水组(NS组)、PTSD+SGB组(SGB组),每组8只。采用条件性恐惧实验建立PTSD模型,检测4组小鼠的恐惧记忆;(2)取3只小鼠,采用伪狂犬病病毒(pseudorabies virus,PRV)逆行示踪观察星状神经节(stellate ganglion,SG)到脑内的神经投射;(3)取24只小鼠,分为空白对照(negative control,NC组)、Sev组、NS组、SGB组,每组6只,采用免疫荧光染色观察蓝斑核(locus coeruleus,LC)去甲肾上腺素(norepinephrine,NE)能神经元(LC^(NE))中c-Fos的表达;(4)取3只小鼠,采用霍乱毒素B亚基(cholera toxin subunit B,CTB)逆行示踪LC到基底外侧杏仁核(basolateral amygdala,BLA)的神经投射;(5)使用化学遗传学和光遗传学方法,借助病毒载体增强神经元活性,观察LC^(NE)-BLA神经环路的化学或光遗传学激活对小鼠恐惧记忆的影响。采用GraphPad Prism 8.0对数据进行单因素方差分析。结果:(1)造模后,NS组小鼠在测试阶段僵立时间百分比[(59.83±6.31)%]与PTSD组[(62.21±7.90)%]和Sev组[(63.61±8.48)%]相比均差异无统计学意义(均P>0.05),SGB组小鼠僵立时间百分比[(47.78±7.02)%]低于NS组(P<0.05)。(2)SG注射PRV病毒示踪结合免疫荧光结果显示,LC^(NE)神经元投射至SG。(3)免疫荧光染色检测小鼠LC^(NE)神经元活动性,结果显示与NC组[(5.52±2.70)%]相比,NS组[(16.75±3.44)%]和Sev组[(14.90±2.73)%]小鼠LC^(NE)神经元中c-Fos表达均明显较高(均P<0.01),SGB组小鼠c-Fos表达[(10.90±3.29)%]较NS组低(P<0.05)。(4)CTB逆行追踪结果显示,LC神经元投射到BLA。(5)SGB小鼠经化学遗传激活后,其僵立时间百分比高于激活前[(67.02±9.49)%,(45.97±7.15)%](P<0.01);SGB组小�Objective To explore the regulation and mechanism of stellate ganglion block(SGB)on conditioned fear memory in post-traumatic stress disorder(PTSD)model mice.Methods The SPF male C57BL/6J mice aged 8-9 weeks were selected and grouped according to the random number table method for the following experiments.(1)A total of 32 mice were divided into PTSD group,PTSD+sevoflurane group(Sev group),PTSD+NS group(NS group)and PTSD+SGB group(SGB group),with 8 mice in each group.The fear memory intensity of mice were assessed by fear conditioned test.(2)The neural projections from the stellate ganglion(SG)to the brain were traced retrograde by pseudorabies virus(PRV)with 3 mice.(3)A total of 24 mice were divided into negative control group(NC group),Sev group,NS group and SGB group,with 6 mice in each group.The expression of c-Fos in locus coeruleus norepinephrine(LC^(NE))neurons of mice was observed by immunofluorescence staining.(4)The neural projections from the locus coeruleus(LC)to the basal lateral amygdala(BLA)were traced retrograde by cholera toxin subunit B(CTB)with 3 mice.(5)Chemogenetic and optogenetic methods were used to enhance the neuronal activity by viral vectors and observe the effects of the LC^(NE)-BLA neural circuit activated by chemogenetic or optogenetic on fear memory in mice.One-way ANOVA was used for statistical analysis by GraphPad Prism 8.0 software.Results(1)After modeling,the percentage of freezing time in NS group((59.83±6.31)%)during test phase showed no statistical difference compared to PTSD group((62.21±7.90)%)and Sev group((63.61±8.48)%)(both P>0.05).And the percentage of freezing time in SGB group((47.78±7.02)%)was lower than that in NS group(P<0.05).(2)The results of injection of PRV virus in SG combined with immunofluorescence showed that LC^(NE) neurons projected to SG.(3)The results of immunofluorescence staining showed that the expressions of c-Fos in LC^(NE) neurons in NS group((16.75±3.44)%)and Sev group((14.90±2.73)%)were significantly higher than that in NC group((5.52±2.7

关 键 词：星状神经节阻滞 创伤后应激障碍 恐惧记忆 蓝斑核 基底外侧杏仁核 小鼠 

分 类 号：R749.5[医药卫生—神经病学与精神病学]