小鼠内耳圆窗基因导入的实验研究

Gene Delivery through Round Window Membrane in Mouse Inner Ear

朱光洁;徐琳;高下;陈杰;丁小琼;麻晓峰;陆玲;秦小明;周函;崔昕燕;

1:东南大学医学院

2:南京大学医学院附属鼓楼医院耳鼻咽喉-头颈外科

摘要
目的研究携带增强型绿色荧光蛋白(enhanced green fluorescent protein,EGFP)腺病毒(Ad-EG-FP)经由小鼠耳后圆窗径路导入内耳的可行性,分析EGFP在耳蜗内的表达特点。方法 19只健康的8~9周龄C57BL/6J雄性小鼠随机分为三组:Ad-EGFP组7只,人工外淋巴液组6只,两组通过耳后切口圆窗径路注射,分别导入Ad-EGFP和人工外淋巴液;空白对照组6只,未予处理。各组均于术前3日和术后7日行听性脑干反应(ABR)检查;术后7日取出耳蜗于荧光显微镜下观察EGFP在内耳的分布并行免疫组化观察EGFP在基底膜的表达。结果 3组动物术前ABR反应阈差异无统计学意义(P>0.05),术后7日,Ad-EGFP组ABR反应阈为62.86±9.94dBSPL,人工外淋巴液组ABR反应阈为60.83±9.70dBSPL,均较术前(37.86±8.59和34.16±8.04dBSPL)及空白对照组(40.83±8.61dBSPL)高(P值均<0.05);空白对照组实验前后ABR反应阈无变化,Ad-EGFP组与人工外淋巴液组术后7天ABR反应阈差异无统计学意义(P值均<0.05)。Ad-EGFP组Ad-EGFP导入后在基底膜上可见EGFP呈广泛表达,人工外淋巴液组和空白对照组基底膜未见荧光表达。结论外源性基因可经内耳圆窗导入并在耳蜗基底膜上广泛表达。
关键词
基因转导;圆窗;腺病毒;小鼠;EGFP
基金项目(Foundation):
国家自然科学基金(30973302);; 江苏省重点人才项目基金(RC2007010);; 南京市医学科技重点项目基金(ZKX06019)资助项目
作者
朱光洁;徐琳;高下;陈杰;丁小琼;麻晓峰;陆玲;秦小明;周函;崔昕燕;
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