用于毛细胞再生研究的噪声性聋动物模型的建立  

A Model of NIHL guinea pig for study of cochlear hair cell regeneration in mammalian

作　　者：陈伟[1,2,3] 郭维维 胡吟燕[1,2] 孙建和[1,2] 于宁[1,2] 李兴启[1,2] 翟所强[1,2] 韩东一[1,2] 杨伟炎[1,2] 杨仕明[1,2] 

机构地区：[1]解放军总医院耳鼻咽喉头颈外科 [2]解放军耳鼻咽喉研究所耳神经生物中心,北京100853 [3]空军总医院耳鼻咽喉头颈外科,北京100142

出　　处：《中华耳科学杂志》2009年第4期327-331,共5页Chinese Journal of Otology

基　　金：国家高技术研究发展计划(863计划)专题项目(2007AA02Z150);国家自然科学基金重点项目(30730040);国家自然科学基金面上项目(30871398;30571017);国家自然科学基金海外青年学者合作基金(30628030)联合资助

摘　　要：目的观察高强度脉冲噪声暴露后豚鼠听功能及耳蜗结构的变化,探讨用于毛细胞再生研究的噪声性聋动物模型的建立方法。方法健康成年白色红目豚鼠50只,雌雄不限,体重250～300g。随机分成2组,正常对照组10只,噪声暴露组40只。给予脉冲噪声(压力峰值为175.0dB SPL,脉宽0.25ms,间隔时间20秒)连续暴露200次。于噪声暴露前及暴露后1周、4周、8周检测听性脑干反应(auditory brainstem respons,ABR),毛细胞计数及耳蜗铺片免疫组化观察耳蜗结构变化。结果高强度脉冲噪声暴露后1周,40只豚鼠中有21只(52.5%)双耳各频率ABR阈值≥95dB SPL。继续观察至噪声暴露后4周及8周,ABR阈值没有恢复,1周、4周、8周各频率ABR阈值比较无统计学差异(P>0.05)。毛细胞计数结果显示,噪声暴露致极重度聋后1周,内毛细胞平均缺失率为91.4%,外毛细胞平均缺失率为97.2%。免疫组化染色分析结果显示,噪声暴露致聋后1周,内、外毛细胞胞核大部分缺失,内毛细胞内侧及外毛细胞外侧的支持细胞的胞核存在。结论高强度脉冲噪声暴露可造成豚鼠极重度感音神经性聋,耳蜗毛细胞广泛缺失且无法自行恢复,而支持细胞大部分存留,是进行毛细胞再生研究的理想动物模型。Objective To study the change of the guinea pig cochlea after high-intensity impulse noise exposure in order to assess its effects on hearing, and to discuss the methods of the noise deafness animal model for cochlea hair cell regeneration research. Methods Fifty adult guinea pigs were used, weight between 250 and 300g, and were divided into two groups in random. Fourty animals were incessant exposed to impulse noise 200 times （Pressure peak was 175.0 dB SPL, and Pulse duration was 0.25 ms, interval time 20s）. For exposed group, 10 were used as control group. Cochlear hair cell counting, Immunohistochemical-dyeing cochlea stretched preparation, and Audtory brainstem response （ABR） thresholds were determined before and 1 week, 4 weeks and 8 weeks after exposed. Results One week after exposed to high-intensity impulse noise, there were 21 of the 40 guinea pigs （52.5%）, with ABR thresholds ≥ 95 dB SPL at every frequency, but not recovered after 8 weeks. No significant change of the ABR thresholds between 1 week, 4 weeks and 8 weeks （P〉 0,05）. Cochlea hair cell counting showed the average miss rate of inner hair cell is 91.4%; the outer hair cell is 97.2% in lweek after noise exposure. Immunohistochemical results showed, that most of the nucleus of the inner hair cell and outer hair cell were missed, but the supporting cell nucleus of the inner hair cell medial and outer hair cell lateral still existed in one week after noise exposure. Conclusion High-intensity impulse noise exposure may cause severe sensorineural deafness, and cochlear hair cells were generally missed and can not recover automatically, but most of the supporting cells were existed. This noise deafness animal model is promise of an ideal animal model for the further study for acoustic hair cell regeneration.

关 键 词：脉冲噪声 豚鼠 耳蜗 毛细胞 ABR 

分 类 号：R322.923[医药卫生—人体解剖和组织胚胎学] R764.433[医药卫生—基础医学]