Nrf2在μ-δ异源二聚体下调EAAT3表达导致大鼠吗啡复吸中的作用  

Role of Nrf2 in morphine relapse caused byμ-δheterodimer-induced down-regulation of EAAT3 expression in rats

作　　者：苗利萍[1] 沈智文[1] 吴毅[1] 张昆[1] 梁建军[1] Miao Liping;Shen Zhiwen;Wu Yi;Zhang Kun;Liang Jianjun(Department of Anesthesiology,Sun Yat-Sen Memorial Hospital,Sun Yat-Sen University,Guangzhou 510280,China)

机构地区：[1]中山大学附属孙逸仙纪念医院麻醉科,广州510280

出　　处：《中华麻醉学杂志》2020年第10期1204-1207,共4页Chinese Journal of Anesthesiology

基　　金：广东省自然科学基金(2016A030313331)。

摘　　要：目的评价核因子E2相关因子2(Nrf2)在μ-δ异源二聚体下调兴奋性氨基酸转运体3(EAAT3)表达导致大鼠吗啡复吸中的作用。方法清洁级健康雄性SD大鼠32只,体重200~240 g,采用随机数字表法分为4组(n=8):对照组(C组)、吗啡戒断组(E组)、吗啡复吸组(R组)和吗啡复吸+干扰质粒组(RI组)。腹腔注射吗啡10 mg/kg,建立大鼠条件性位置偏爱(CPP)模型;停止给药使CPP逐渐消退;再次腹腔注射小剂量吗啡5 mg/kg诱导已消退的CPP恢复,记录伴药箱停留时间。CPP模型建立成功后RI组大鼠侧脑室注射μ-δ异源二聚体干扰质粒5μl。采用高效液相色谱法测定海马谷氨酸含量,采用Western blot法检测海马内Nrf2和EAAT3的表达,采用RT-PCR法检测海马内Nrf2 mRNA的表达,采用蛋白免疫共沉淀法检测μ-δ异源二聚体与Nrf2相互作用情况。结果与C组比较,R组和RI伴药箱停留时间延长,海马谷氨酸含量和μ-δ异源二聚体/Nrf2比值升高,E组和RI组海马Nrf2和EAAT3表达上调,E组、R组和RI组海马Nrf2 mRNA表达上调(P<0.05);与E组比较,R组和RI组伴药箱停留时间延长,海马谷氨酸含量和μ-δ异源二聚体/Nrf2比值升高,Nrf2和EAAT3表达下调(P<0.05),海马Nrf2 mRNA表达差异无统计意义(P>0.05);与R组比较,RI组伴药箱停留时间缩短,海马谷氨酸含量和μ-δ异源二聚体/Nrf2比值降低,海马Nrf2和EAAT3表达上调(P<0.05),海马Nrf2 mRNA表达差异无统计意义(P>0.05)。结论μ-δ异源二聚体形成后结合Nrf2,导致海马Nrf2含量减少,引起EAAT3表达下调,从而导致大鼠吗啡复吸的形成。Objective To evaluate the role of Nrf2 in morphine relapse caused byμ-δheterodimer-induced down-regulation of excitatory amino acid transportor 3(EAAT3)expression in rats.Methods Thirty-two clean-grade healthy male Sprague-Dawley rats,weighing 200-240 g,were divided into 4 groups(n=8 each)using a random number table method:control group(group C),morphine withdrawal group(group E),morphine relapse group(group R),and morphine relapse plus interference plasmid group(group RI).Morphine 10 mg/kg was intraperitoneally injected to establish conditioned place preference(CPP)model;administration was stopped to make CPP gradually subside;a small dose of morphine 5 mg/kg was intraperitoneally injected again to induce the relapse of the subsided CPP,and the time spent in drug-paired box was recorded.In group RI,μ-δheterodimer interference plasmid 5μl was injected into the lateral ventricle after CPP model was successfully established.The content of glutamate in hippocampus was determined by high-performance liquid chromatography,and the expression of Nrf2 and EAAT3 was detected by Western blot,the expression of Nrf2 mRNA was detected by real-time polymerase chain reaction,and the interaction betweenμ-δheterodimer and Nrf2 was detected by protein immunoprecipitation method.Results Compared with group C,the time spent in drug-paired box was significantly prolonged,and the glutamate content andμ-δheterodimer/Nrf2 ratio were increased in R and RI groups,the expression of Nrf2 and EAAT3 was significantly up-regulated in E and RI groups,and the expression ofNrf2 mRNA was significantly up-regulated in E,R and RI groups(P<0.05).Compared with group E,the time spent in drug-paired box was significantly prolonged,the glutamate content andμ-δheterodimer/Nrf2 ratio were increased,the expression of Nrf2 and EAAT3 was down-regulated,and no significant change was found in Nrf2 mRNA expression in R and RI groups(P>0.05).Compared with group R,the time spent in drug-paired box was significantly shortened,the glutamate content andμ-

关 键 词：受体 阿片样 吗啡依赖 兴奋性氨基酸转运体3 NF-E2相关因子2 海马 异源二聚体 

分 类 号：R614[医药卫生—麻醉学]