听力学及言语疾病杂志
聽力學及言語疾病雜誌
은역학급언어질병잡지
JOURNAL OF AUDIOLOGY AND SPEECH PATHOLOGY
2014年
3期
272-276
,共5页
刘渊%尹时华%黄训健%周琦%舒竞铖%韦顺莲
劉淵%尹時華%黃訓健%週琦%舒競鋮%韋順蓮
류연%윤시화%황훈건%주기%서경성%위순련
艾芬地尔%水杨酸钠%耳蜗%圆窗膜%螺旋神经节
艾芬地爾%水楊痠鈉%耳蝸%圓窗膜%螺鏇神經節
애분지이%수양산납%이와%원창막%라선신경절
Ifenprodil%Sodium salicylate%Cochlear%Round window membrane%Spiral ganglion neu-ron
目的:通过圆窗龛局部给予特异性N R2B受体阻滞剂艾芬地尔,探讨其拮抗水杨酸钠致豚鼠螺旋神经节细胞损伤的作用。方法将ABR阈值小于40 dB SPL的健康花色豚鼠48只随机分为4组,每组12只,Ⅰ组为空白对照组;Ⅱ组为人工外淋巴液(artificial perilymph ,APL)组,左耳圆窗龛注人 APL 60μl;Ⅲ组为水杨酸钠组,左耳圆窗龛注入A PL 60μl ,腹腔注射水杨酸钠;IV组为艾芬地尔组,左耳圆窗龛注入溶于 A PL的艾芬地尔(10μmol/L )60μl后,腹腔注射水杨酸钠。水杨酸钠注射量为400 mg · kg -1· d-1,连续注射7天。停药并行ABR测试后将动物处死,取出左侧耳蜗,每组随机取6只动物进行免疫组织化学染色观察耳蜗螺旋神经节(SGN ) caspase-3表达情况,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。结果给药后,I、II、III、IV 组动物左耳ABR反应阈分别为31.67±5.16、33.33±5.17、64.17±7.36、49.17±5.85 dB SPL ;免疫组织化学染色显示,Ⅰ、Ⅱ组螺旋神经节细胞中caspase-3的表达量差异无统计学意义,Ⅲ、IV组螺旋神经节细胞中caspase-3的表达量较Ⅰ、Ⅱ组显著增高(P<0.01),IV组螺旋神经节细胞中caspase-3的表达量较Ⅲ组明显降低(P<0.01)。TUNEL法检测结果示,Ⅰ、Ⅱ组SGN细胞凋亡率无显著差异,Ⅲ、IV组SGN细胞凋亡率较Ⅰ、Ⅱ组显著增高(P<0.01), IV组SGN细胞凋亡率较Ⅲ组明显降低(P<0.01)。结论耳蜗局部特异性阻断NR2B受体可以拮抗水杨酸钠致豚鼠螺旋神经节细胞的损伤。
目的:通過圓窗龕跼部給予特異性N R2B受體阻滯劑艾芬地爾,探討其拮抗水楊痠鈉緻豚鼠螺鏇神經節細胞損傷的作用。方法將ABR閾值小于40 dB SPL的健康花色豚鼠48隻隨機分為4組,每組12隻,Ⅰ組為空白對照組;Ⅱ組為人工外淋巴液(artificial perilymph ,APL)組,左耳圓窗龕註人 APL 60μl;Ⅲ組為水楊痠鈉組,左耳圓窗龕註入A PL 60μl ,腹腔註射水楊痠鈉;IV組為艾芬地爾組,左耳圓窗龕註入溶于 A PL的艾芬地爾(10μmol/L )60μl後,腹腔註射水楊痠鈉。水楊痠鈉註射量為400 mg · kg -1· d-1,連續註射7天。停藥併行ABR測試後將動物處死,取齣左側耳蝸,每組隨機取6隻動物進行免疫組織化學染色觀察耳蝸螺鏇神經節(SGN ) caspase-3錶達情況,6隻行原位末耑轉移酶標記技術(terminal deoxynucleotidyl transferase -mediated dUTP -biotin nick end labeling assay ,TUNEL)法檢測螺鏇神經節細胞凋亡率。結果給藥後,I、II、III、IV 組動物左耳ABR反應閾分彆為31.67±5.16、33.33±5.17、64.17±7.36、49.17±5.85 dB SPL ;免疫組織化學染色顯示,Ⅰ、Ⅱ組螺鏇神經節細胞中caspase-3的錶達量差異無統計學意義,Ⅲ、IV組螺鏇神經節細胞中caspase-3的錶達量較Ⅰ、Ⅱ組顯著增高(P<0.01),IV組螺鏇神經節細胞中caspase-3的錶達量較Ⅲ組明顯降低(P<0.01)。TUNEL法檢測結果示,Ⅰ、Ⅱ組SGN細胞凋亡率無顯著差異,Ⅲ、IV組SGN細胞凋亡率較Ⅰ、Ⅱ組顯著增高(P<0.01), IV組SGN細胞凋亡率較Ⅲ組明顯降低(P<0.01)。結論耳蝸跼部特異性阻斷NR2B受體可以拮抗水楊痠鈉緻豚鼠螺鏇神經節細胞的損傷。
목적:통과원창감국부급여특이성N R2B수체조체제애분지이,탐토기길항수양산납치돈서라선신경절세포손상적작용。방법장ABR역치소우40 dB SPL적건강화색돈서48지수궤분위4조,매조12지,Ⅰ조위공백대조조;Ⅱ조위인공외림파액(artificial perilymph ,APL)조,좌이원창감주인 APL 60μl;Ⅲ조위수양산납조,좌이원창감주입A PL 60μl ,복강주사수양산납;IV조위애분지이조,좌이원창감주입용우 A PL적애분지이(10μmol/L )60μl후,복강주사수양산납。수양산납주사량위400 mg · kg -1· d-1,련속주사7천。정약병행ABR측시후장동물처사,취출좌측이와,매조수궤취6지동물진행면역조직화학염색관찰이와라선신경절(SGN ) caspase-3표체정황,6지행원위말단전이매표기기술(terminal deoxynucleotidyl transferase -mediated dUTP -biotin nick end labeling assay ,TUNEL)법검측라선신경절세포조망솔。결과급약후,I、II、III、IV 조동물좌이ABR반응역분별위31.67±5.16、33.33±5.17、64.17±7.36、49.17±5.85 dB SPL ;면역조직화학염색현시,Ⅰ、Ⅱ조라선신경절세포중caspase-3적표체량차이무통계학의의,Ⅲ、IV조라선신경절세포중caspase-3적표체량교Ⅰ、Ⅱ조현저증고(P<0.01),IV조라선신경절세포중caspase-3적표체량교Ⅲ조명현강저(P<0.01)。TUNEL법검측결과시,Ⅰ、Ⅱ조SGN세포조망솔무현저차이,Ⅲ、IV조SGN세포조망솔교Ⅰ、Ⅱ조현저증고(P<0.01), IV조SGN세포조망솔교Ⅲ조명현강저(P<0.01)。결론이와국부특이성조단NR2B수체가이길항수양산납치돈서라선신경절세포적손상。
Objective To investigate whether blocking NR2B receptor can reverse the process of cytotoxicity to spiral ganglion neurons induced by sodium salicylate in guinea pig by applying ifenprodil (a NR2B antagonist) at the round window niche .Methods Sixty healthy guinea pigs provided by the experimental animal center of Guangxi medical university were randomly and evenly divided into a control group (Group I ,no treatment) ,an APL group (Group II ,60μl APL directly applied to the round window ) ,a sodium salicylate group (Group III ,60 μl APL di-rectly applied to the round window and then be given intraperitoneal sodium salicylate injection ) ,and an ifenprodil group (Group IV ,60μl of 10μmol/l ifenprodil in APL directly applied to the round window and then be given intra-peritoneal sodium salicylate injection ) .Sodium salicylate was given at 400 mg · kg -1 · d-1 for 7 days .Auditory brainstem responses (ABRs) were recorded 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 TUNEL technique .Results Before salicylate administration ,the ABR threshold was less than 40 dB SPL in all animals .After salicylate ad-ministration ,the ABR threshold was 33 .33 ± 5 .17 dB SPL in Group II ,64 .17 ± 7 .36 dB SPL in Group III and 49 .17 ± 5 .85 dB SPL in Group IV ,in contrast to 31 .67 ± 5 .16 dB SPL in Group I (controls) .The caspase -3 ex-pression was not changed obviously in Group I and Group II ,but was significantly changed in Group III and Group IV (P<0 .01) .The caspase-3 expression appeared to be decreased in Group IV compared to those in Groups III (P<0 .05) ,but still increased compared to those of in Group I and II (P<0 .05) .The apoptosis index among spiral ganglion neurons in Groups III and IV increased significantly compared to those of in Group I and II (P<0 .001) .It was however ,lower in Group IV than in Group III (P<0 .01) .Conclusion Blocking NR2B receptor with specificity can reverse the process of cytotoxicity to spiral ganglion neurons induced by sodium salicylate in guinea pig .