中华耳科学杂志
中華耳科學雜誌
중화이과학잡지
CHINESE JOURNAL OF OTOLOGY
2012年
4期
521-527
,共7页
周琦%刘渊%唐安洲%谭颂华%黄训健%尹时华
週琦%劉淵%唐安洲%譚頌華%黃訓健%尹時華
주기%류연%당안주%담송화%황훈건%윤시화
美金刚%阿米卡星%耳蜗%圆窗膜%螺旋神经节
美金剛%阿米卡星%耳蝸%圓窗膜%螺鏇神經節
미금강%아미잡성%이와%원창막%라선신경절
memantine%amikacin%cochlear%round window membrane%spiral ganglion neuron
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5 mg/ml,60μl后,肌肉注射阿米卡星).阿米卡星注射量为400 mg/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 transfer?ase-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±3.34 dB SPL ,Ⅱ组:90.42±5.42 dBSPL,Ⅲ组:91.17±5.37 dB SPL ,IV组:78.75±6.78 dB SPL.免疫组织化学染色显示,Ⅱ组、Ⅲ组动物耳蜗螺旋神经节caspase 3表达较Ⅰ组升高(P<0.01).IV组动物耳蜗螺旋节caspase 3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caspase 3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肌肉注射阿米卡星减少.美金刚经圆窗龛给药对阿米卡星螺旋神经节细胞毒性有一定拮抗作用.
目的通過圓窗龕跼部給藥,探討美金剛對阿米卡星緻豚鼠螺鏇神經節細胞毒性的拮抗作用.方法將聽性腦榦反應(auditory brainstem response, ABR)檢測結果小于40dBSPL的花色豚鼠共60隻,隨機分為4組,每組15隻.Ⅰ組:空白對照組;Ⅱ組:單純阿米卡星給藥組(肌肉註射阿米卡星);Ⅲ組:人工外淋巴液(artificial perilymph , APL)+阿米卡星給藥組(左耳圓窗龕註人APL60μl後,肌肉註射阿米卡星);IV組:美金剛+阿米卡星給藥組(左耳圓窗龕註入溶于APL的美金剛5 mg/ml,60μl後,肌肉註射阿米卡星).阿米卡星註射量為400 mg/kg/d,連續註射5天,停藥14天後,每組動物行ABR閾值檢測,將動物處死,取齣每組動物左耳蝸.每組隨機取6隻通過免疫組織化學染色觀察耳蝸螺鏇神經節caspase3錶達情況.剩下6隻行原位末耑轉移酶標記技術(terminal deoxynucleotidyl transfer?ase-mediated dUTP-biotin nick end labeling assay,TUNEL)法檢測螺鏇神經節細胞凋亡率.結果給藥前,各組動物ABR閾值檢測結果均小于40dBSPL.給藥後,各組動物左耳ABR閾值結果為:Ⅰ組:37.08±3.34 dB SPL ,Ⅱ組:90.42±5.42 dBSPL,Ⅲ組:91.17±5.37 dB SPL ,IV組:78.75±6.78 dB SPL.免疫組織化學染色顯示,Ⅱ組、Ⅲ組動物耳蝸螺鏇神經節caspase 3錶達較Ⅰ組升高(P<0.01).IV組動物耳蝸螺鏇節caspase 3錶達較Ⅱ組、Ⅲ組降低(P<0.05),較Ⅰ組升高(P<0.01).TUNEL法檢測螺鏇神經節細胞凋亡率:Ⅱ組、Ⅲ組動物耳蝸螺鏇神經節細胞凋亡率較Ⅰ組明顯升高(P<0.001).IV組動物耳蝸螺鏇神經節凋亡率較Ⅱ組、Ⅲ組降低(P<0.01),較Ⅰ組升高(P<0.001).結論(1)美金剛經圓窗龕跼部給藥後肌肉註射阿米卡星,豚鼠ABR閾值低于單純肌肉註射阿米卡星ABR閾值.(2)美金剛經圓窗龕跼部給藥後肌肉註射阿米卡星,螺鏇神經節caspase 3錶達較單純肌肉註射阿米卡星減少.(3)美金剛經圓窗龕跼部給藥後肌肉註射阿米卡星,螺鏇神經節凋亡較單純肌肉註射阿米卡星減少.美金剛經圓窗龕給藥對阿米卡星螺鏇神經節細胞毒性有一定拮抗作用.
목적통과원창감국부급약,탐토미금강대아미잡성치돈서라선신경절세포독성적길항작용.방법장은성뇌간반응(auditory brainstem response, ABR)검측결과소우40dBSPL적화색돈서공60지,수궤분위4조,매조15지.Ⅰ조:공백대조조;Ⅱ조:단순아미잡성급약조(기육주사아미잡성);Ⅲ조:인공외림파액(artificial perilymph , APL)+아미잡성급약조(좌이원창감주인APL60μl후,기육주사아미잡성);IV조:미금강+아미잡성급약조(좌이원창감주입용우APL적미금강5 mg/ml,60μl후,기육주사아미잡성).아미잡성주사량위400 mg/kg/d,련속주사5천,정약14천후,매조동물행ABR역치검측,장동물처사,취출매조동물좌이와.매조수궤취6지통과면역조직화학염색관찰이와라선신경절caspase3표체정황.잉하6지행원위말단전이매표기기술(terminal deoxynucleotidyl transfer?ase-mediated dUTP-biotin nick end labeling assay,TUNEL)법검측라선신경절세포조망솔.결과급약전,각조동물ABR역치검측결과균소우40dBSPL.급약후,각조동물좌이ABR역치결과위:Ⅰ조:37.08±3.34 dB SPL ,Ⅱ조:90.42±5.42 dBSPL,Ⅲ조:91.17±5.37 dB SPL ,IV조:78.75±6.78 dB SPL.면역조직화학염색현시,Ⅱ조、Ⅲ조동물이와라선신경절caspase 3표체교Ⅰ조승고(P<0.01).IV조동물이와라선절caspase 3표체교Ⅱ조、Ⅲ조강저(P<0.05),교Ⅰ조승고(P<0.01).TUNEL법검측라선신경절세포조망솔:Ⅱ조、Ⅲ조동물이와라선신경절세포조망솔교Ⅰ조명현승고(P<0.001).IV조동물이와라선신경절조망솔교Ⅱ조、Ⅲ조강저(P<0.01),교Ⅰ조승고(P<0.001).결론(1)미금강경원창감국부급약후기육주사아미잡성,돈서ABR역치저우단순기육주사아미잡성ABR역치.(2)미금강경원창감국부급약후기육주사아미잡성,라선신경절caspase 3표체교단순기육주사아미잡성감소.(3)미금강경원창감국부급약후기육주사아미잡성,라선신경절조망교단순기육주사아미잡성감소.미금강경원창감급약대아미잡성라선신경절세포독성유일정길항작용.
Objective To investigate the antagonism of memantine toward amikacin cytotoxicity 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, amikacin 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 amikacin group (Group IV, 60 μl of 5 mg/ml memantine in APL directly applied to the round window followed by intramuscular amikacin injection). Amikacin was given at 400 mg/kg/d for 5 days. Auditory brainstem responses (ABRs) were recorded 14 days later before animal sacrifice by decapitation. 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 37.08±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 IIand III (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 IIand III (P<0.01). Conclusion(1)Local application of memantine reduces ABR threshold shift caused by amikacin administration.(2)Local application,of memantine also decreases caspase 3 expression, as well as the apoptosis index, in spiral ganglion neurons in response to amikacin administration.(3)Memantine appears to antagonize the amikacin cytotoxicity to spiral ganglion neurons.
