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内耳注射药物建立新生鼠耳聋动物模型

A Hearing Loss Model in Mouse Neonate by Direct Drug Delivery to the Cochlea

胡凌翔;吴皓;石复辛;

1:上海交通大学耳科学研究所内耳发育与再生实验室

2:上海交通大学医学院附属新华医院耳鼻咽喉头颈外科

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摘要

目的通过纳升级微量注射泵内耳注射给药的实验方法,建立新生鼠耳聋动物模型。方法 42只出生第二天新生鼠,分别以注射速度分组:单纯穿刺组(对照组,6只,仅穿刺耳蜗底回侧壁,不注射液体)、蒸馏水1组(H2O 20组,4只,20nl/min)、蒸馏水2组(H2O 60组,3只,60nl/min)及药物浓度分组(新霉素各组给药速度为60nl/min):新霉素1组(Neo1组,4只,1μmol/ml)、新霉素2组(Neo10组,4只,10μmol/ml)、新霉素3组Neo 50组,5只,50μmol/ml)、新霉素4组(Neo 100组,8只,100μmol/ml)及新霉素5组(Neo 50 1组,8只,50μmol/ml)。各组均采用耳蜗底回微创打孔技术,利用纳升级微量注射泵按上述各组用药、剂量及给药速度进行内耳药物注射。术后1天(Neo 50 1组)及3天(其余各组)采用基底膜免疫荧光染色观察内耳毛细胞及支持细胞的形态改变。结果术后新生鼠存活好,伤口恢复优良。耳蜗毛细胞机械性损伤与注射速度呈正相关,术后3天,H2O 20组和H2O 60组注射区域内、外毛细胞略减少,差异无统计学意义(P>0.05)。毛细胞损伤与新霉素浓度呈正相关,与对照组比较,术后3天Neo1组的外毛细胞数量及Neo10、50、100组内、外毛细胞数量均明显减少(P<0.05)。与H2O 60组比较,术后3天Neo50及Neo100组内、外毛细胞数量均明显减少(P<0.05)。新霉素50μmol/ml注射后1天(Neo 50 1组)与3天(Neo 50组)内、外毛细胞数量差异无统计学意义(P>0.05)。新霉素50μmol/ml注射后3天免疫荧光染色示支持细胞形态良好。结论利用纳升级微量注射泵给药方式注射50μmol/ml新霉素200纳升至内耳,可建立新生鼠耳聋动物模型,具有毛细胞死亡率高,致聋效果显著的优点;而支持细胞保留较好,且动物存活率高。

关键词

内耳注射;耳聋模型;新霉素

基金项目(Foundation):

国家自然科学基金面上项目(81170924)

作者

胡凌翔;吴皓;石复辛;

参考文献

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胡凌翔吴皓石复辛