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腺病毒携带EGFP基因经完整圆窗膜转导豚鼠耳蜗的实验研究

ISSN号：1672-2922
期刊名称：《中华耳科学杂志》
时间：0
分类：R339.16[医药卫生—人体生理学;医药卫生—基础医学] R349.34[医药卫生—基础医学]
作者机构：[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,北京100853, [2]美国Creighton大学医学院生物医学系,美国NE68175
相关基金：本课题由国家自然科学基金海外青年学者合作基金（No.30628030）,国家自然科学基金面上项目（No.30000189.No.30571017）,北京市自然科学基金（No.7042061）资助.

作者：陈伟[1], 杨仕明[1], 郭维[1], 胡吟燕[1], 孙建和[1], 韩东一[1], 翟所强[1], 杨伟炎[1], 何志洲

关键词：基因转导, 圆窗膜, 腺病毒, 豚鼠, EGFP, Gene transfer, Round window membrane, Adenovirus, Guinea pig, EGFP

中文摘要：

目的研究腺病毒携带目的基因经完整圆窗膜途径耳蜗转导的可行性及安全性，为内耳基因治疗提供实验基础和理论依据。方法20只白色红目豚鼠，术前及术后分别行听性脑干反应（ABR）检查。实验组（12只）以重组腺病毒携带的增强型绿色荧光蛋白基因（enhancedgreenfluorescentprotein，EGFP），对照组（8只）以人工外淋巴液注入豚鼠圆窗龛内。分别于术后5天、14天取双侧耳蜗标本做基底膜铺片，耳蜗冰冻切片观察。结果于圆窗龛内注入腺病毒携带目的基因的转导方法对听力无明显影响。转染耳蜗及对侧耳蜗内目的基因呈广泛表达。5天组表达产物最高，14天组逐渐降低。对照组耳蜗未见EPFP表达。结论于圆窗龛内注入腺病毒携带目的基因转导的方法对耳蜗无明显毒害作用，且能够将目的基因成功转导至双侧耳蜗组织并广泛表达。

英文摘要：

Objective To investigate delivery of recombinant abenoviruses-erhanced green fluorescent protein （Ad- EGFP） into the guinea pig cochlea through an intact round window membrane（RWM）, and to assess the side-effect of the management. Methods Twenty adult guinea pigs were used for the study： 12 were implanted with Ad-EGFP in the bony groove of the round window niche, and 8 were implanted with artificial perilymphatic fluid. Auditory brainstem response （ABR） thresholds were determined in all animals before and 5 days after surgery. On postoperative day 5 and day 14, the animals were sacrificed and the surface preparation of cochleae was observed. Results ABR measurement showed that the operation had no deleterious effect on hearing. Bright green fluorescence in the cochleae was observed in Ad-EGFP groups. Gene expression was at a high level 5 days after operation and reduced 14 days later. However, control groups were free of fluorescence. Conclusion The technique of transgenic delivery into the inner ear through an intact RWM is feasible, atraumatic and effective.

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