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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(enhanced green fluorescent protein,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。
Objective To study the feasibility and safety of adenoviral transduction of the target gene in cochlear transduction through a complete round window membrane and to provide experimental basis and theoretical basis for gene therapy of inner ear. Methods Twenty white - necked guinea pigs were examined with auditory brainstem response (ABR) before and after operation. The experimental group (12 mice) was treated with recombinant adenovirus carrying enhanced green fluorescent protein (EGFP) and the control group (8 mice) with artificial perilymph into guinea pig round window niche. Bilateral cochlear specimens were taken at 5 days and 14 days after operation to make the basement membrane and the cochlear frozen sections were observed. Results In the round window niche, the transduction method of adenovirus carrying the target gene had no significant effect on hearing. Transfection of the cochlear and contralateral cochlear gene was widely expressed. The highest expression product was found on the 5th day and gradually decreased on the 14th day. There was no EPFP expression in the control group. Conclusion The method of transplanting adenovirus into target gene in round window niche has no obvious toxic effect on the cochlea, and can successfully transduce the target gene into bilateral cochlear tissues and widely expressed.