异氟醚对大鼠海马组织小窝蛋白家族表达的影响  

A study of the expression and the significance of caveolin family in rat hippocampus upon exposing to multiple doses of isoflurane

作　　者：蔡琳[1] 隋大鸣[1] 付海钰 査鹏 古小平[1] 巩固[1] CAI Lin;SUI Daming;FU Haiyu;ZHA Peng;GU Xiaoping;GONG Gu(Department of Anesthesiology,General Hospital of Chengdu Military Region of PLA,Chengdu 610083,China)

机构地区：[1]成都军区总医院麻醉科,610083

出　　处：《中国神经免疫学和神经病学杂志》2018年第3期170-175,共6页Chinese Journal of Neuroimmunology and Neurology

基　　金：2016年军队后勤科研项目(CCD16J001)

摘　　要：目的探索异氟醚对大鼠海马区小窝蛋白家族(Cav-1、Cav-2、Cav-3)表达的影响。方法将SD大鼠随机分为对照组、异氟醚1组(ISO1)、异氟醚2组(ISO2)和异氟醚3组(ISO3),每组10只,对照组吸入40%氧气,ISO1吸入1.2%异氟醚,ISO2吸入1.8%异氟醚,ISO3吸入2.4%异氟醚,分别处理2h。Morris水迷宫实验检测各组大鼠苏醒后第1~5天逃避潜伏期及空间探索能力。利用qRT-PCR和Western blot检测大鼠海马组织Cav、脑源性神经营养因子(BDNF)、细胞外信号调节蛋白激酶(ERK1/2)、酪氨酸受体激酶B(TrkB)的表达。构建腺病毒载体Ad-Cav-1及对照Ad-GFP转染海马神经元,经异氟醚处理后采用噻唑蓝(MTT)法检测原代海马神经元存活率。结果大鼠麻醉苏醒后2~4d,ISO1、ISO2和ISO3组逃避潜伏期长于对照组(均P<0.05),而各组大鼠1d和5d逃避潜伏期无统计学差异(均P>0.05);各组大鼠第5天空间探索时间比较无统计学差异(F=2.057,P=0.154)。与对照组比较,ISO1、ISO2和ISO3组大鼠Cav-1、Cav-2、Cav-3mRNA和蛋白表达水平降低(均P<0.05)。ISO1、ISO2和ISO3组大鼠BDNF、TrkB、磷酸化ERK1/2(p-ERK1/2)表达水平亦明显低于对照组(均P<0.05)。Ad-Cav-1转染促进海马神经元Cav-1的表达(P<0.05),并提高异氟醚处理后神经元存活(P<0.05)。结论异氟醚可降低大鼠海马组织Cav的表达,过表达Cav-1则具有拮抗异氟醚的作用。Objective To explore the effect of isoflurane on caveolins（Cav-1,Cav-2 and Cav-3）expression and the role of caveolins in hippocampal neurons. Methods Sprague-Dawly rats were randomly allocated into a control group,isoflurane group 1（ISO1）,isoflurane group 2（ISO2）,and isoflurane group 3（ISO3）,with 10 mice in each group. Rats in the control group were administrated with 40% oxygen,ISO1 group done with 1.2% isoflurane,ISO2 group done with 1.8% isoflurane,and ISO3 done with 2.4% isoflurane,2 h for each group. Escape latency at 1-5 d upon waking up and spatial learning ability of these rats were determined by Morris water maze test. At 24 h after waking up,rat hippocampal tissues were prepared. The expressions of caveolins,brain derived neurotrophic factor（BDNF）,extracellular signal-regulated protein kinase（ERK1/2）,and tyrosine receptor kinase B（TrkB）in hippocampal tissues were examined by qRT-PCR and immunoblotting. Adenovirus Ad-Cav-1 and its control Ad-GFP were constructed and transfected into primary hippocampal neurons. After isoflurane exposure,neuron viability was detected by MTT assay. Results Upon waking up from 2 d to 4 d,the escape latencies of the ISO1,ISO2 and ISO3 groups were markedly prolonged compared with that of the control group （all P 〈0.05）;at 1 d and 5 d,the escape latencies among all the groups varied insignificantly（all P 〉0.05）;at 5 d,no significant difference was found in the spatial probe time among all the groups（ F =2.057, P =0.154）. Compared with the controls,mRNA and protein levels of Cav-1,Cav-2,and Cav-3 of rats in the ISO1,ISO2 and ISO3 groups were significantly reduced（all P 〈0.05）. The expressions of BDNF,TrkB and phosphorylated ERK1/2（p-ERK1/2）also decreased in the ISO1,ISO2 and ISO3 groups compared with the control group（all P 〈0.05）. Infection of Ad-Cav-1 promoted Cav-1 expression（ P 〈0.05）and cell viability in neurons treated with isoflurane（ P 〈0.05）. Conclusions Isoflurane decreases caveolins expressio

关 键 词：异氟醚 小窝蛋白 海马神经元 脑源性神经营养因子 细胞外调节蛋白激酶 

分 类 号：R747.9[医药卫生—神经病学与精神病学] R971.2[医药卫生—临床医学]