美金刚拮抗阿米卡星对豚鼠螺旋神经节细胞毒性的实验研究  

Antagonistic Effects of Memantine against Amikacin Cytotoxicity to Spiral Ganglion Neurons in Guinea Pigs

作　　者：周琦[1] 刘渊[1] 唐安洲[1] 谭颂华[1] 黄训健[1] 尹时华[1] 

机构地区：[1]广西医科大学第一附属医院耳鼻咽喉头颈外科,广西530021

出　　处：《中华耳科学杂志》2012年第4期521-527,共7页Chinese Journal of Otology

基　　金：国家自然科学基金(项目基金编号:30860312);广西自然科学基金(项目基金编号:桂科自0728134)

摘　　要：目的通过圆窗龛局部给药,探讨美金刚对阿米卡星致豚鼠螺旋神经节细胞毒性的拮抗作用。方法将听性脑干反应(auditory brainstem response,ABR)检测结果小于40dBSPL的花色豚鼠共60只,随机分为4组,每组15只。Ⅰ组:空白对照组;Ⅱ组:单纯阿米卡星给药组(肌肉注射阿米卡星);Ⅲ组:人工外淋巴液(artificial perilymph,APL)+阿米卡星给药组(左耳圆窗龛注人APL60μl后,肌肉注射阿米卡星);IV组:美金刚+阿米卡星给药组(左耳圆窗龛注入溶于APL的美金刚5mg/ml,60μl后,肌肉注射阿米卡星)。阿米卡星注射量为400mg/kg/d,连续注射5天,停药14天后,每组动物行ABR阈值检测,将动物处死,取出每组动物左耳蜗。每组随机取6只通过免疫组织化学染色观察耳蜗螺旋神经节caspase3表达情况。剩下6只行原位末端转移酶标记技术(terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay,TUNEL)法检测螺旋神经节细胞凋亡率。结果给药前,各组动物ABR阈值检测结果均小于40dBSPL。给药后,各组动物左耳ABR阈值结果为:Ⅰ组:37.08±3.34dB SPL,Ⅱ组:90.42±5.42dBSPL,Ⅲ组:91.17±5.37dB SPL,IV组:78.75±6.78dB SPL。免疫组织化学染色显示,Ⅱ组、Ⅲ组动物耳蜗螺旋神经节caspase3表达较Ⅰ组升高(P<0.01)。IV组动物耳蜗螺旋节caspase3表达较Ⅱ组、Ⅲ组降低(P<0.05),较Ⅰ组升高(P<0.01)。TUNEL法检测螺旋神经节细胞凋亡率:Ⅱ组、Ⅲ组动物耳蜗螺旋神经节细胞凋亡率较Ⅰ组明显升高(P<0.001)。IV组动物耳蜗螺旋神经节凋亡率较Ⅱ组、Ⅲ组降低(P<0.01),较Ⅰ组升高(P<0.001)。结论 (1)美金刚经圆窗龛局部给药后肌肉注射阿米卡星,豚鼠ABR阈值低于单纯肌肉注射阿米卡星ABR阈值。(2)美金刚经圆窗龛局部给药后肌肉注射阿米卡星,螺旋神经节caspase3表达较单纯肌肉注射阿米卡星减少。(3)美金刚经圆窗龛局部给药后肌肉注射阿米卡星,螺旋神经节凋亡较单纯�Objective To investigate the antagonism of memantine toward amikaein eytotoxieity to spiral ganglion neu- rons in guinea pigs when it is applied at the round window niche. Methods Sixty healthy guinea pigs provided by the Experi- mental Animal Center of Guangxi Medical University were randomly and evenly divided into a control group.（Group I）, an ami- kacin group （Group II, amikaein given intramuscularly）, an APL plus amikacin group （Group III, 60 μl APL directly applied to the round window followed by intramuscular amikacin injection）, and a memantine plus amikaein group （Group IV, 60 μl of 5 mg/ml memantine in APL directly applied to the round window followed by intramuscular amikacin injection）. Amikaein was given at 400 mg/kg/d for 5 days. Auditory brainstem responses （ABRs） were recorded 14 days later before animal sacrifice bydecapitation. The left cochlea was removed and prepared for detection of caspase 3 expression in spiral ganglion neurons via immunohistochemistry. From each group, 6 cochleae were used to test apoptosis index in spiral ganglion neurons using the TU- NEL technique. Results Before amikacin administration ,the ABR threshold was less than 40 dB SPL in all animals. After ami- kacin administration, the ABR threshold was 90.42±5.42 dB SPL in Group II, ,91.17±5.37 dB SPL in Group III and 78.75±6.78 dB SPL in Group IV, in contrast to 57.08±3.34 dB SPL in Group I （controls）. The caspase 3 expression was not obvious in group I, but significantly visible in Groups IIand III （P〈0.01）. The caspase 3 expression appeared to be decreased in Group IV compared to that in Groups Iland IIl （P〈0.05）, but still increased compared to that in Group I （P〈0.05）. The apoptosis index among spiral ganglion neurons in Groups II, III and IV increased significantly compared to that in Group I （P〈0.001）. It is how- ever, lower in Group IV than in Groups Iland III （P〈O.OI）. Conclusion （1） Local application of memantine reduces ABR threshold shift caused by

关 键 词：美金刚 阿米卡星 耳蜗 圆窗膜 螺旋神经节 

分 类 号：R764.4[医药卫生—耳鼻咽喉科]