CAJ | 학술논문

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C57BU6J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 fluoreseent 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.结论经腹侧听泡外人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.
목적 연구선병독휴대목적 기인경복측은포외입로전도이와고계저전적가행성급목적 기인적표체특점.위내이기인치료제공실험기출화이론의거.방법 16지건강5주령C57BU6J소서,선병독조10지,이중조선병독휴대유Hath-1화증강형록색형광단백기인(enhanced green fluoreseent protein,EGFP),인공외림파액조6지이인공외림파액,경복측은포외입로도입이와고계저전.분별우술후제7천분별행은성뇌간반응(ABR)검사후취쌍측이와표본주기저막포편、이와빙동절편관찰기인적표체.결과경복측은포외입로전도이와고계저전적전도방법 대은력영향교소.선병독조이와내목적 기인정엄범표체.대조조이와미견형광표체.결론 경복측은포외인로전도이와고계저전적전도방법 대은력영향교소,차능구장목적 기인성공전도지이와조직병엄범표체.
Objective To assess the feasibility of adenoviral vector-mediated gene transfer into the scala tympani via a ventral approach microinjection without opening the tympanic bulla in mice. Methods Ad -Hath -7-EGFP was delivered into the scala tympani in ten C57BIV6J 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 microinjection into the scala tympani without opening the bulla is an effective means of gene transfer into the cochlea with minimal disturbance to auditory mechanisms.