七氟醚暴露通过抑制终纹床核中的胞外信号调节激酶缓解幼年小鼠创伤后应激障碍  

Sevoflurane exposure alleviate posttraumatic stress disorder by inhibiting extracellular signal-regulated kinase activation in the bed nucleus of stria terminalis in juvenile mice

作　　者：周凯意 胡家祺 吴亚男 梁杰贤 雷黎明 罗昕 钟锋[2] 王晟 Zhou Kaiyi;Hu Jiaqi;Wu Yanan;Liang Jiexian;Lei Liming;Luo Xin;Zhong Feng;Wang Sheng(School of Medicine,South China University of Technology,Guangzhou 510006,China;Department of Anesthesiology,Guangdong Provincial People's Hospital(Guangdong Academy of Medical Sciences),Southern Medical University,Guangzhou 510080,China;Southern Medical University,the Great Bay Areea Center for Brain Research,Guangzhou 510515,China;Anaesthesia Center of Beijing Anzhen Hospital,Capital Medical University,Bejing 100029,China)

机构地区：[1]华南理工大学医学院,广州510006 [2]南方医科大学附属广东省人民医院(广东省医学科学院)麻醉科,广州510080 [3]南方医科大学粤港澳大湾区脑科学和类脑研究中心,广州510515 [4]首都医科大学附属安贞医院麻醉中心,北京100029

出　　处：《国际麻醉学与复苏杂志》2024年第11期1135-1140,共6页International Journal of Anesthesiology and Resuscitation

基　　金：国家自然科学基金地区科学基金项目(82160157);国家自然科学基金青年科学基金项目(82101258);国家自然科学基金面上项目(32371043);广东省“珠江人才计划-青年拔尖人才”项目(2021QN02Y169);广东省医学科学技术研究基金项目(B2021387);广州市基础研究计划基础与应用基础研究项目(1611)。

摘　　要：目的探讨终纹床核(BNST)中的磷酸化胞外信号调节激酶(p-ERK)在七氟醚暴露缓解幼年小鼠创伤后应激障碍的调控作用。方法取3周龄SPF级雄性C57/BL小鼠43只,按随机数字表法分为6组:对照组(C组,11只)、七氟醚组(S组,12只)、对照+溶媒组(C1组,5只)、对照+ERK抑制剂组(C2组,5只)、七氟醚+溶媒组(S1组,5只)、七氟醚+ERK激动剂组(S2组,5只)。S组、S1组、S2组小鼠连续3d暴露于3%七氟醚+30%氧气,每天2h;C组、C1组、C2组小鼠连续3d暴露于30%氧气,每天2h。S组、S1组、S2组小鼠七氟醚干预后1d间断给予足底电击,建立应激增强恐惧学习(SEFL)模型;C组、S组小鼠在第2天进行创伤场景测试,观察小鼠僵直行为,计算僵直行为时间百分比(T1);6组小鼠在第3天进行电击-新场景配对,观察小鼠僵直行为,计算小鼠基础僵直行为时间百分比(T2);6组小鼠在第4天进行敏化测试,观察小鼠僵直行为,计算小鼠敏化测试僵直行为时间百分比(T3)。敏化测试前30min,C2组小鼠腹腔注射ERK抑制剂U012610mg/kg,S2组小鼠腹腔注射ERK激动剂Honokiol10mg/kg,C1组、S1组小鼠腹腔注射生理盐水500μl,C组和S组不做处理。敏化测试结束后处死小鼠,取脑组织,采用免疫组织化学方法检测6组小鼠BNST中p-ERK水平,采用免疫荧光方法检测C组和S组BNST中p-ERK与囊泡型谷氨酸转运体2(VGLUT2)的共标情况。结果与C组比较,S组小鼠T1和T3均降低(均P<0.05),T2差异无统计学意义(P>0.05);与C1组比较C2组小鼠T3降低(P<0.05),T2差异无统计学意义(P>0.05);与S1组比较,S2组T3明显升高(P<0.05),T2差异无统计学意义(P>0.05)。与C组比较,S组小鼠BNST中p-ERK阳性细胞面积减少(P<0.05);与C1组比较,C2组小鼠BNST中p-ERK阳性细胞面积减少(P<0.05);与S1组比较,S2组小鼠BNST中p-ERK阳性细胞面积增加(P<0.05);与C2组比较,S2组小鼠BNST中p-ERK阳性细胞面积增加(P<0.05)。C组小鼠在BNST激活的ERK阳性神经元中有较多VGLUTObjective To investigate the regulatory role of phosphorylated extracellular signal-regulated kinase(p-ERK)in the bed nucleus of stria terminalis(BNST)in alleviating post-traumatic stress disorder in juvenile mice with sevoflurane exposure.Methods SPF male C57/BL mice aged 3 weeks were randomly divided into 6 groups:control group(group C,n=11),sevoflurane group(group S,n=12),control+Vehicle group(group Cl,n=5),control+extracellular signal-regulated kinase(ERK)inhibitor group(group C2,n=5),sevoflurane+Vehicle group(group S1,n=5),sevoflurane+ERK agonist group(group S2,n=5).Mice in group S,group S1,and group S2 were exposed to 3%sevoflurane+30%oxygen for 3 consecutive days for 2 h per day,and mice in group C,group C1,and group C2 were exposed to 30%oxygen for 3 consecutive days for 2 h per day.The mice in group S,group S1,group S2 were treated with sevoflurane on the lst day,and the model of stress-enhanced fear learning was established.The mice in group C and group S were subjected to trauma scene test on the second day to observe the rigidity behavior of the mice,the percentage of stiff behavior time(T1)was calculated,and six groups of mice were paired with electric shock-new scene on the 3rd day to observe the stiff behavior and the percentage of time of basal stiffness(T2)was calculated;the 6 groups of mice underwent sensitization test on day 4,observed the stff behavior of mice,and calculated the percentage of time(T3)of mouse sensitization test stiff behavior.Thirty minutes before the sensitization test,mice in group C2 were intraperitoneally injected with ERK inhibitor UO12610 mg/kg,mice in group S2 were intraperitoneally injected with ERK agonist Honokiol 10 mg/kg,and mice in group Cl and group S1 were intraperitoneally injected with 500μl normal saline,group C and group S were not treated.The expression of p-ERK in bed nucleus of stria terminalis(BNST)of 6 groups of mice was detected by immunohistochemistry,the co-labeling of p-ERK and vesicular glutamate transporter 2(VGLUT 2)in BNST was detected by immun

关 键 词：七氟醚 胞外信号调节激酶 创伤后应激障碍 终纹床核 幼年小鼠 

分 类 号：R614[医药卫生—麻醉学]