不同固定液对大鼠耳蜗一氧化氮合酶表达及透射电镜观察的影响  

The Effects of Different Fixative on Transmission Electron Microscopy Examination and Induced Nitric Oxide Synthase Expression in Rat

作　　者：陈小婉[1] 江英[2] 仇志强[1] 张小兵[1] 

机构地区：[1]兰州大学第一医院耳鼻咽喉-头颈外科,兰州730000 [2]兰州大学

出　　处：《听力学及言语疾病杂志》2016年第1期58-61,共4页Journal of Audiology and Speech Pathology

基　　金：兰州大学第一医院青年基金资助(ldyyynqn201005)

摘　　要：目的探寻用于观察大鼠耳蜗诱生型一氧化氮合酶(induced nitric oxide synthase,iNOS)表达及透射电镜检查最适合的固定液配比,简化实验手段。方法将正常大鼠6只(12耳,正常组)和庆大霉素耳中毒模型大鼠6只(12耳,模型组)各分为A、B两组(每组各3只,6耳),A组给予2.5%多聚甲醛-2.5%戊二醛、B组给予4%多聚甲醛-0.5%戊二醛混合液心内灌注耳蜗固定处死,然后进行耳蜗免疫组化染色及透射电镜观察,比较两组耳蜗的染色和电镜固定效果。结果正常组与模型组大鼠的耳蜗外毛细胞、螺旋神经节等均有iNOS阳性表达;但正常组及模型组的A组耳蜗组织免疫组化染色有轻度溶解,背景染色不佳,B组耳蜗组织细胞染色清晰,对比度及透明度好,背景无明显着色。正常组及模型组中A、B两组电镜下耳蜗细胞器等微结构清晰。结论 4%多聚甲醛-0.5%戊二醛混合液心内灌注的方法既能较好地固定大鼠耳蜗细胞器的结构,也能较好的保留iNOS抗原。Objective To find a more suitable fixative for examining transmission electron microscopy（TEM）and induced nitric oxide synthase（iNOS）measure.Methods A total of 12 adult male rats（24ears）with normal hearing were used in this study and randomly divided into control group（6rats,12ears）and gentamicin group（6rats,12ears）.Three rats in each group was fixed with 4%paraform-0.5% glutaraldehyde mixture,the another 3rats were fixed with 2.5%paraform-2.5% glutaraldehyde mixture.The cochleae were examined by transmission electron microscopy（TEM）to observe ultrastructural changes.In addition,iNOS expression was measured.Results The positive staining of iNOS was observed in the cochlea in all groups.The control groups were weakly positive,while the gentamicin groups were strongly positive.Those tissue sections showed cell dissolved,uneven dyeing,poor background staining which were fixed with 2.5% paraform-2.5% glutaraldehyde mixture.While there were no significant differences between different fixed groups in TEM results.Conclusion Our resucts indicate that 4%paraform-0.5% glutaraldehyde mixture is better than 2.5%paraform-2.5% glutaraldehyde mixture for fixing rat by heart perfusing for TEM examine and iNOS expression measure.

关 键 词：大鼠 诱生型一氧化氮合酶 透射电镜 固定液 庆大霉素 

分 类 号：R764.3[医药卫生—耳鼻咽喉科]