Neuroglobin 基因体内转染对水杨酸钠给药后小鼠下丘核神经元听反应的影响(英文)  

Effects of neuroglobin gene transfer in vivo on hearing response properties of neurons in the inferior colliculus in mice after administration of sodium salicylate

作　　者：尹时华[1] 龚树生[1] 鄢开胜[1] 李穗[1] 陈沛[1] 陈广理[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院耳鼻咽喉科,武汉430022

出　　处：《生理学报》2005年第4期529-536,共8页Acta Physiologica Sinica

基　　金：This work was supported by the National Award for Outstanding Junior Faculty Member issued by Ministry of Education of China.

摘　　要：实验以48只成年健康昆明小鼠为实验对象,研究GeneJamer转染试剂介导的neuroglobin(NGB)基因体内转染对水杨酸钠给药后小鼠下丘核区听反应的影响。实验分4组,每组12只。A1组:对照组1(阴性对照,将GeneJamer转染试剂6μl和pEGFP-C12μg混合后注入下丘核脑区);A2组:对照组2[阳性对照,将GeneJamer转染试剂(6μl)和pEGFP-NGB(质粒载体pEGFP-C1与NGB基因全编码序列构建的重组子2μg)混合后注入下丘核脑区];B组:水杨酸钠给药组(450mg/kg·d-1)+pEGFP-C1;C组:水杨酸钠(450mg/kg.d-1)+pEGFP-NGB组。以直接注射法将GeneJamer转染试剂和重组质粒pEGFP-NGB混合后注入小鼠下丘核区。采用RT-PCR和Westernblot技术检测小鼠下丘核区NGBmRNA和蛋白的表达;采用细胞外记录技术,研究小鼠下丘核区神经元在水杨酸钠给药后转染重组质粒pEGFP-NGB对强度-发放率函数(刺激声强与实验鼠下丘核区神经元在接受声刺激所产生的电发放的关系曲线)、强度-潜伏期函数(刺激声强与实验鼠下丘核区神经元在接受声刺激至产生电发放潜伏期之间的关系曲线)和频率调谐曲线(实验鼠下丘核区神经元在各个频率纯音刺激下起反应的阈值绘制的曲线)的影响。实验观察到:(1)经GeneJamer转染试剂介导NGB基因可有效地转染小鼠下丘核区脑组织并得到表达。(2)水杨酸钠给药后神经元的强度-发放率函数曲线升高。对照组A1、A2各项指标进行比较均无统计学意义。对照组A1、A2和水杨酸钠+pEGFP-NGB组神经元的强度-发放率函数以非单调型(随刺激强度增加时,发放率表现为先降后升呈“V”形或“U”形)为主,分别占74.6%、72.2%和59.3%,水杨酸钠给药组以不规则型强度-发放率函数为主,占47%,与对照组A1、A2和水杨酸钠+pEGFP-NGB组比较,有显著性差异(P<0.01、P<0.01、P<0.05)。(3)水杨酸钠给药后神经元的强度-潜伏期函数曲线降低。对照组A1、A2各项指标进行比较均无统The effects of neuroglobin （NGB） gene transfer in vivo mediated by GeneJamer on the hearing response properties of the inferior colliculus （IC） neurons in mice after administration of sodium salicylate were studied. Forty-eight Kunming mice were divided into 4 groups （n=12 in each group）： Group A1 （negative control）; Group A2 （positive control）; Group B, sodium salicylate （450 mg/kg every day） ＋ pEGFP-C1; Group C, sodium salicylate （450 mg/kg every day） ＋ pEGFP-NGB. The GeneJamer and pEGFP-NGB were mixed and injected into IC neurons in mice. The expression of NGB mRNA and protein of IC neurons in mice was detected by using RT-PCR and Western blot methods. The intensity-rate functions, intensity-latency functions and frequency-turning curves in IC neurons were recorded by extracellular electrophysiological recording techniques and the effects of pEGFP-NGB transfer following injection of sodium salicylate on them were studied. It was found that： （1） The GeneJamer-mediated pEGFP-NGB could be effectively transferred into the IC brain tissues in mice and NGB could be expressed intensively. （2） The intensity-rate functions of IC neurons were raised after administration of sodium salicylate. The non-monotonic styles of intensity-rate functions in groups A 1, A2 and C were accounted for 74.6 %, 72.2 %, 59.3 %, respectively, and the function in group B for 47%. There were significant differences between group B and groups A1, A2 or C （P〈0.01, P〈0.01, P〈0.05）. （3） The intensity-latency functions in 1C neurons were reduced after administration of sodium salicylate. The non-monotonic styles of intensity-latency functions in groups A1, A2 and C were accounted for 3.2 %, 5.1% and 21%, respectively, and that in group B for 45.5 %. There were significant differences between group B and groups A1, A2 or C （P〈0.01, P〈0.01, P〈0.05, respectively）. （4） The frequency-turning curves in groups A1 and A2 were sharpened. In 72 acoustic neurons recorded in the group B,

关 键 词：水杨酸钠 小鼠 下丘核 基本特性 脑红素 基因治疗法 

分 类 号：Q426[生物学—神经生物学] R338.4[生物学—生理学]