摘要
目的探索Math1基因导入大鼠前庭简便有效的方法和途径,为前庭功能障碍基因治疗的相关研究提供参考。方法将20只成年Wistar大鼠分为缺失E1、E3基因片段且构建有Math1基因和增强型绿色荧光蛋白报告基因的复制缺陷型腺病毒(adnovirus-Math1-enhanced green fluorescence protein,Ad-Math1-EGFP)鼓阶导入组和前庭阶导入组,Ad-Math1-EGFP导入组大鼠在右耳通过耳蜗底转鼓阶或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。在导入3天、7天后分别将动物处死,进行GFP表达观察。结果导入Ad-Math1-EGFP3天后,前庭阶导入组大鼠的前庭终末器官及耳蜗均出现明显的GFP阳性表达;而鼓阶导入组的表达则局限于耳蜗,7天后仍未见前庭终末器官的GFP阳性表达。结论耳蜗底转前庭阶打孔可以作为Math1基因导入大鼠前庭简便有效的途径。
Objective To explore the best way of Math1 gene transfer into vestibule and provide the base data for gene therapy for vestibular dysfunction. Methods Twenty mature wistar rats were divided into normal control group, the group of adenovirus （E1, E3-Deleted and carried Math1 gene and enhanced green fluorescent protein report gene, Ad- Math1-EGFP） scala tympani transfering and scala vestibuli transfering group. Right ears of the Ad-Mathl-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 tympani or scala vestibuli of cochlear basal turn. Animals were killed at 3 and 7 days respectively after treatment, and then were observed expression of GFP. Results All the vestibular end organs were transfeeted after treatment 3 days in the group of Ad-Math1-EGFP scala vestibuli transfer, but could not observe the transfection in Ad- Math1-EGFP scala tympani transfering group until 7 days after treatment, exception of cochlea. Conclusion The cochleotomy of basal turn through scale vestibular can be served as simple and effective way of transfering Mathl gene into vestibule.
出处
《中华耳科学杂志》
CSCD
2009年第4期342-346,共5页
Chinese Journal of Otology
基金
国家自然科学基金(面上项目30871398
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
关键词
基因导入
前庭
大鼠
腺病毒
Vestibule
Gene transfer
Rat
Adenovirus
