腹侧听泡外入路小鼠耳蜗鼓阶基因导入的实验研究  

Intact bulla approach for gene transfer into the scala tympani in mice

作　　者：徐延军[1] 杨仕明[1] 胡吟燕[1] 翟所强[1] 孙建和[1] 徐金操[1] 候昭晖[1] 申卫东[1] 于宁[1] 韩东一[1] 

机构地区：[1]解放军总医院耳鼻咽喉头颈外科解放军耳鼻咽喉研究所耳神经生物中心,北京100853

出　　处：《中华耳科学杂志》2009年第1期79-84,共6页Chinese Journal of Otology

基　　金：国家自然科学基金重点项目(30730040);国家863计划专题项目(2007AA02Z150);国家自然科学基金海外青年学者合作基金(30628030);国家自然科学基金面上项目(30571017)

摘　　要：目的研究腺病毒携带目的基因经腹侧听泡外入路转导耳蜗鼓阶底转的可行性及目的基因的表达特点,为内耳基因治疗提供实验基础和理论依据。方法16只健康5周龄C57BL/6J小鼠,腺病毒组10只,以重组腺病毒携带有Hath-1和增强型绿色荧光蛋白基因(enhanced green fluorescent protein,EGFP),人工外淋巴液组6只以人工外淋巴液,经腹侧听泡外入路导入耳蜗鼓阶底转。分别于术后第7天分别行听性脑干反应(ABR)检查后取双侧耳蜗标本做基底膜铺片、耳蜗冰冻切片观察基因的表达。结果经腹侧听泡外入路转导耳蜗鼓阶底转的转导方法对听力影响较小。腺病毒组耳蜗内目的基因呈广泛表达。对照组耳蜗未见荧光表达。结论经腹侧听泡外入路转导耳蜗鼓阶底转的转导方法对听力影响较小,且能够将目的基因成功转导至耳蜗组织并广泛表达。Objective To assess the feasibility of adenoviral vector-mediated gene transfer into the scala tympani via a ventral approach microinjeetion without opening the tympanic bulla in mice. Methods Ad-Hath-I-EGFP was delivered into the scala tympani in ten C57BL/6J mice via a ventral approach without opening the bulla, while another six mice were injected with artificial perilymphatic fluid through the same approach as the control. Auditory brainstem response （ABR） thresholds were determined in all animals 7 days after injection. The animals were then sacrificed for cochlear surface preparation and section specimens. Results ABRs indicate minimal hearing disturbance by the injection procedure. Ad-EGFP expression in the cochlea was demonstrated by bright green fluorescence in animals injected with Ad-Hath-1-EGFP, while control animals showed no presence of fluorescence. Conclusion The results indicate that ventral approach microinjeetion into the scala tympani without opening the bulla is an effective means of gene transfer into the cochlea with minimal disturbance to auditory mechanisms.

关 键 词：基因转导 鼓阶 腺病毒 小鼠 EGFP 

分 类 号：R322.923[医药卫生—人体解剖和组织胚胎学] R349.6[医药卫生—基础医学]