耳蜗TRPC6基因导入治疗大鼠噪音性聋的初步研究  

The study of TRPC6 gene transfer into cochlea to cure the rats’noise-induced hearing lose

作　　者：单良 徐雨笠 李玎 童军 Shan Liang;Xu Yuli;Li Ding;Tong Jun(Department of Otolaryngology,Shanghai Fourth People’s Hospital Afiliated to Tongji University School of Medicine,Shanghai 200434,China)

机构地区：[1]同济大学附属上海市第四人民医院耳鼻咽喉科,上海200434

出　　处：《遵义医科大学学报》2021年第3期285-289,共5页Journal of Zunyi Medical University

基　　金：上海市卫生和计划生育委员会科研课题计划(NO:201640183)。

摘　　要：目的探讨TRPC6(瞬时受体电势C通道蛋白6)过表达慢病毒(PHBCMV-TRPC-EGFP)导入SD大鼠耳蜗后,能否转染毛细胞,促进听力修复。方法21只健康大鼠,随机分为3组:TRPC6组,空病毒组,对照组。噪音暴露后,TRPC6组和空病毒组,于噪声暴露后1d经圆窗膜耳蜗分别注入PHBCMV-TRPC-EGFP和荧光表达空病毒。全部3组大鼠于噪音暴露后3、10、20 d行脑干电位(ABR)检测,噪声暴露后20 d行耳蜗铺片,观察耳蜗TRPC6的转染状况及毛细胞变化。结果噪音暴露前及后3 d,3组大鼠ABR值差异无统计学意义。噪音暴露后10 d,ABR值差异有统计学意义:其中TRPC6组与空病毒和对照组之间ABR阈值差异有显著性(P=0.01,P=0.00)。噪音暴露后20 d,3组ABR值差异有统计学意义(P=0.00),其中TRPC6组与空病毒和对照组之间差异均有显著性(P=0.00,P=0.00)。噪音暴露后20 d,TRPC6组和空病毒组大鼠耳蜗铺片在荧光显微镜下和GFP激光共聚焦显微镜下观察,可见荧光阳性表达毛细胞,对照组耳蜗铺片未见毛细胞荧光表达。TRPC6组较空病毒组毛细胞缺失较轻。结论TRPC6过表达慢病毒导入大鼠耳蜗后,能成功转染大鼠毛细胞,促进其修复以治疗噪音性耳聋。Objective To explore whether the microinjection of TRPC6 overexpressing lentiviral vector carrying EGFP(PHBCMV-TRPC-EGFP)into the cochlea can transfer the gene into hair cells,then help the SD rats’injury hair cells recovery and cure the noise-induced hearing lose.Methods Twenty one healthy rats were randomly divided into three groups:the TRPC6 Group,the Empty Virus Group and the Control Group.All rats were noise exposed.The TRPC6 Group and the Empty Virus Group were microinjected with PHBCMV-TRPC-EGFP and empty lentiviral vector carrying EGFP into cochlea through round window respectivly 1 day after noise exposed.The ABR test are made in all three groups at 3,10,20 days after noise exposed.Twenty days after noise exposed,Cochlear preparations were collected from all rats to exam the results of the change of hair cells via GFP expression.Results There was no significant difference in the ABR test results among three groups before and 3 days after noise exposure.And there were significant differences in that 10,20 days after noise exposure respectivly.the TRPC6 group had a lower ABR threshold compare with the Empty Virus Group(P=0.01,P=0.00,10 days after)and Control Group(P=0.00,P=0.00,20 days after).The hair cells could be seen on cochlear preparetion via GFP expression in the two microinjected groups.The TRPC6 Group show a milder hair cells’damage than the Empty Virus Group.Conclusion The TRPC6 gene can be introduced into the cochlea through a round window microinjection and expressed in the rat’s hair cells.Therefore it can reduce the damage of the cochlear hair cells and promote the of recovery of noise-induced hearing loss.

关 键 词：TRPC6 基因转导 耳蜗 噪音性聋 

分 类 号：R764.3[医药卫生—耳鼻咽喉科]