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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 study the feasibility of transfection of the target gene by adenoviruses through the ventral peritoneal bulb and the expression characteristics of the target genes in order to provide the experimental basis and theoretical basis for the gene therapy of the inner ear. Methods Sixteen healthy 5-week-old C57BL / 6J mice and 10 adenovirus groups were infected with recombinant adenovirus carrying Hath-1 and enhanced green fluorescent protein (EGFP) Only artificial perilymph, through the ventral to listen to the outer tube into the cochlear drum at the end of rotation. On the 7th day after operation, bilateral cochlear specimens were taken from the auditory brainstem response (ABR) to make the basement membrane and the cochlear frozen sections to observe the gene expression. The results of the transvenous extraperitoneal approach transduced cochlear scleral buckling transduction method has little effect on hearing. Adenovirus group of cochlear gene was widely expressed. The control group showed no fluorescence expression in the cochlea. Conclusion The transduction methods of transdotransduction of cochlear basal ganglia via ventral telangiectasia have less influence on hearing, and can transduce the gene of interest into cochlea and express it extensively.