Math1/神经生长因子β基因共转染对噪声性听力损伤豚鼠耳蜗毛细胞的保护作用  

Protective effect of Math1/NGFβ gene transfer on hair cells in guinea pigs with noise-induce hearing damage

作　　者：戴尉君 张健 吴建 

机构地区：[1]第二军医大学第二附属医院耳鼻咽喉科,上海200003

出　　处：《中华航海医学与高气压医学杂志》2013年第6期365-368,388,共5页Chinese Journal of Nautical Medicine and Hyperbaric Medicine

基　　金：全军医药卫生“十二五”科研基金课题（CWS11J128）;全军医药卫生“十二五”科研基金重点课题（BWS11C035）

摘　　要：目的 观察Math1基因（mouse atonal homologue 1 gene,Math1）及神经生长因子β基因（nerve growth factor beta gene,NGFβ）共转染对噪声性听力损伤豚鼠耳蜗毛细胞的保护作用.方法 选用35只白色纯种豚鼠,制备豚鼠稳态噪声耳聋模型（110 dB SPL）,噪声暴露前后分别行听觉脑干电位（auditory brainstem response,ABR）检测,选择噪声暴露前后听阈阈移大于60 dB SPL的豚鼠.采用数字表法随机分为4组,其中A组10只,为双基因组（导入目的基因Ad-Math 1/NGFβ）;B组10只,为Math1组（导入目的基因Ad-Math1）;C组10只,为NGF组（导入目的基因Ad-NGFβ）;D组5只,为对照组（导入空病毒）.基因转染后,进行ABR反应阈测定、免疫荧光染色检测基因蛋白表达,并用电镜扫描观察耳蜗毛细胞.结果 基因转染后1周,Ad-Math1/NGFβ、Ad-Math1、Ad-NGFβ在豚鼠耳蜗内成功转染,耳蜗各回膜性组织均有表达,强度基本相等;A组双基因转染豚鼠ABR反应阈恢复显著快于B、C组单基因转染豚鼠ABR反应阈;转染后2周,A组豚鼠ABR[（37.6±2.8） dB SPL]已基本恢复正常,而B、C组豚鼠ABR[（45.3±2.5） dB SPL及（47.5±3.1） dB SPL]没有恢复正常,D组豚鼠ABR没有恢复[（75.7±3.4）dB SPL],A组与其他3组比较,差异均有统计学意义（P＜0.01）.扫描电镜示：噪声暴露后可见各组豚鼠耳蜗毛细胞静纤毛融合及缺失;基因转染后1周,A组豚鼠耳蜗可见新生毛细胞出现,其他组豚鼠耳蜗未见新生毛细胞;转染后2周,A组豚鼠耳蜗新生毛细胞数量增加,B、C、D组豚鼠耳蜗未见新生耳蜗毛细胞.结论 Math 1/NGFβ基因共转染在噪声损伤后豚鼠耳蜗中能高效表达,对噪声性听力损伤的保护作用明显优于单基因转染.Objective To observe the protective effect of Mathl/NGFβ gene transfer on hair cells in guinea pigs with stable noise-induced hearing damage.Methods Thirty-five healthy white guinea pigs were used to develop the model of noise-induced hearing damage （110 dB SPL）.Measurement of auditory brainstem response （ABR） was made both before and after noise exposure.Rats with hearing threshold shifts greater than 60 dB were chosen for the experiment.Then,the animals were randomly divided into 4 groups：the Ad-Math1/NGFβ group or group A with 10 animals,the Ad-Math1 group or group B with 10 animals,the Ad-NGFβ group or group C also with 10 animals,and the control group or group D with 5 animals.Following gene transfer,ABR threshold shifts were measured,expressions of genes were detected with immunofluorescent staining method and morphologic changes in cochlear hair cells were observed with scanning electron microscopy.Results One week after gene transfer,the said gene was successfully expressed in each turn of thc cochlea forthe animals of the first 3 groups,with the intensity of expression being almost identical.ABR threshold shifts for the animals of group A recovered significantly faster than those in the animals of group B and group C （P 〈0.01）.Two weeks after gene transfer,ABR threshold shifts for the animals of group A returned almost to normal level[（37.6 ±2.8） dB SPL],while threshold shifts for the animals of group B and group C failed to return to normal levels [（45.3 ± 2.5） dB SPL and （47.5 ± 3.1） dB SPL],and threshold shifts for the animals of group D didn＇ t return to normal level [（75.7 ± 3.4） dB SPL],and statistical significance could be seen between the 4 groups （P 〈 0.01）.Scanning electron microscopy indicated that hair cell stereocilium integration or deficiency could be noted in all the animal groups,following noise exposure.One week after gene transfer,new hair cells appeared in the cochlea of the animals of group A,while new hair cells were not observed in the

关 键 词：Math1基因 神经生长因子β基因 噪声 毛细胞 基因治疗 

分 类 号：R764[医药卫生—耳鼻咽喉科]