腺伴随病毒介导ADNF-9基因转染对豚鼠卡那霉素致聋的治疗作用  

Treatment of adeno-associated virus mediated ADNF-9 gene transfection on guinea pig cochlea deafened by kanamycin

作　　者：景阳[1] 郑国玺[2] 祝康[2] 刘晖[1] 张文[1] 

机构地区：[1]陕西省人民医院耳鼻咽喉科,陕西西安710068 [2]西安交通大学第二附属医院耳鼻咽喉科,陕西西安710004

出　　处：《江苏大学学报（医学版）》2015年第5期369-372,共4页Journal of Jiangsu University：Medicine Edition

基　　金：国家自然科学基金资助项目(81271057);陕西省科技研究发展计划资助项目(2013K12-12-04)

摘　　要：目的:观察包装重组的腺伴随病毒(adeno-associated virus,AAV)r AAV-NT4-ADNF-9对卡拉霉素致聋豚鼠的治疗作用。方法:选用耳廓反射和听性脑干反应(auditory brainstem response,ABR)正常的白色红目豚鼠,在大腿内侧肌肉注射卡那霉素400 mg/(kg·d),连续注射6 d,制备耳聋模型。在全身麻醉的条件下,从豚鼠右耳背侧进路,打开听泡,用微量注射器由圆窗注入r AAV-NT4-ADNF-9 10μL,14 d后取术侧听泡。进行耳蜗毛细胞计数和耳蜗毛细胞的硬铺片,通过相差显微镜观察内耳毛细胞的结构。结果:r AAV-NT4-ADNF-9组治疗后ABR阈值较治疗前明显下降(P<0.05),相差显微镜观察到耳蜗基膜完整,外毛细胞呈点状缺失。而其余3组治疗前后ABR阈值无明显变化,人工淋巴液组与空载体组治疗后毛细胞呈片状缺失,空白对照组则无毛细胞缺失。ADNF组取景框内毛细胞平均数明显高于人工淋巴液对照组及空载体组(P<0.05)。结论:用r AAV-NT4-ADNF-9转染卡那霉素致聋豚鼠耳蜗组织,可促进耳蜗毛细胞修复和再生。Objective:To observe the effect of rAAV-NT4-ADNF-9 in treating ototoxicity associated with aminoglycoside antibiotics.Methods:The guinea pigs were used,and its auvicle reflex and auditory brain-stem response(ABR)were normal.The ototoxic animal model of guinea pigs were made by the method of hypodemic injection of Kanamycin(400 mg/kg).In animals under general anesthesia,the guinea pigs were cut off the skin of ringt ear sulcus posterior and opened acoustic capsule,then injected the rAAV-NT4-AD-NF-9 10 μL from fossula rotunda.After 14 days,the right acoustic capsule were dislodged.The cochlear were stained with AgNO3 .The cochlear hair cells were observed by contrast phase microscope.Results:Taking the group for rAAV-NT4-ADNF-9 as example,the threshold ABR decreased remarkably after treat-ment(P <0.05).Through the phase contrast microscope the cochlear basilar membrane was complete and the outer hair cells had dot deletion.The thresholds of ABR had no notable differences for the other three groups before and after treatment.Taking comparison between the artificial endolymph control group and empty vector group,through the phase contrast microscope after treatment,hair cells showed a patchy loss in the former group and there was no such phenomenon in the latter group.The average number of hair cells observed in the group of ADNF was significantly higher than that of the artificial endolymph control group and empty vector group(P <0.05).Conclusion:rAAV-NT4-ADNF-9 is able to be transfected into cochle-ar,and to express secretory NT4-ADNF-9 peptide,which preventing hair cells from impairment.

关 键 词：基因治疗 感音神经性耳聋 活性依赖性神经营养因子-9 耳蜗毛细胞 腺伴随病毒 

分 类 号：R764.431[医药卫生—耳鼻咽喉科]