复制缺陷型腺病毒为载体的Math1基因内耳应用安全性研究  

Evaluation of the Safety of Adenovirus-Math1 Administration to Inner Ear

作　　者：徐金操[1] 胡吟燕[2] 徐延军[2] 孙建和[2] 黄德亮[2] 杨仕明[2] 

机构地区：[1]解放军第二炮兵总医院耳鼻咽喉头颈外科,北京100853 [2]解放军总医院耳鼻咽喉-头颈外科解放军耳鼻咽喉研究所

出　　处：《听力学及言语疾病杂志》2010年第1期35-39,共5页Journal of Audiology and Speech Pathology

基　　金：国家自然科学基金(面上项目30571017;30000189);海外青年合作基金(30628030);北京自然科学基金(7042061);国家863计划(2007AA02Z150);国家自然科学基金重点项目(30730040)资助

摘　　要：目的研究以复制缺陷型腺病毒为载体的Math1基因内耳应用的安全性。方法将10只成年Wistar大鼠分为正常对照组和缺失E1、E3基因片段且构建有Math1基因和绿色荧光蛋白报告基因的复制缺陷型腺病毒(adenovirus-Math1-enhanced green fluorescence protein,Ad-Math1-EGFP)前庭阶导入组(实验组),每组5只,实验组大鼠在右耳通过耳蜗底回前庭阶打孔的方法导入物理滴度为2.1×1011v.p/ml的上述腺病毒5μl,对照组大鼠不做任何处理。7天后对动物进行颈髓硬膜外短声诱发电位(click-evoked potentials on the surface of the cervical dura mater,CDM-CEP)、听性脑干反应(ABR)阈值检测和游泳试验,评价前庭和耳蜗功能,然后将动物处死进行组织形态学观察。结果Ad-Math1-EGFP导入7天后,实验组大鼠前庭及耳蜗的毛细胞及纤毛均未见破坏,形态正常。短声刺激下,对照组大鼠CDM-CEP的阈值为85±3.54dBSPL,ABR阈值为37±4.47dBSPL;实验组大鼠CDM-CEP的阈值为89±6.52dBSPL,ABR阈值为40±3.54dBSPL;对照组大鼠的游泳时间为4.0±0.71s,实验组为5.0±0.71s,两组之间比较差异均无统计学意义。结论携带Math1基因的复制缺陷型腺病毒对前庭和耳蜗毛细胞是安全的,可以作为基因导入的理想载体。Objective To evaluate the safety of Ad-Math1 administration to inner ear and provide the base data for vestibular dysfunction gene therapy. Methods Ten mature Wistar rats were divided into normal control group（srats） and adenovirus（E1,E3-Deleted and carried math1 and enhanced green fluorescent protein report gene,Ad-Math1-EGFP）scala vestibuli transfer group（5 rats）. Right ears of the Ad-Math1-EGFP transfering group rats were deliveried 5ul Ad-Math1-EGFP（physical tite 2.1 1011v.p./ml） into cochleas through the way of drilling scala vestibuli of cochlear basal turn. As a control,the normal group received nothing to inner ear. In order to estimate functional condition of vestibule and cochlea,the click-evoked potentials on the surface of the cervical dura mater（CDM-CEP）,auditory brain stem response（ABR） and swimming time were recorded in all rats at 7 days after treatment,and then histologic and morphologic observation were carried out after animals were sacrificed. Results All animals＇ morphologic observation showed that inner ear hair cells were normal after transfer.Seven days after transfer,the swimming time was 4.0±0.71 s in normal control group and 5.0±0.71 s in scala vestibuli delivery group. The threshold of CDM-CEP and ABR were 85±3.54 dB SPL and 37±4.47 dB SPL in normal control group,and 89±6.52 dB SPL and 40±3.54 dB SPL in Ad-Math1-EGFP scala vestibuli delivery group,respectively. There was no significant difference existed between control group and Ad-Math1-EGFP scala vestibuli delivery group. Conclusion The Ad（E1,E3-Deleted ）is safe for vestibular and cochlea hair cells and can be used as an ideal vector of gene transfer.

关 键 词：Math1基因 安全性 腺病毒 内耳 

分 类 号：R764.3[医药卫生—耳鼻咽喉科]