新霉素致损后听觉上皮TFB1M、线粒体、毛细胞凋亡相关性研究杨大志.docVIP

新霉素致损后听觉上皮TFB1M、线粒体、毛细胞凋亡相关性研究 杨大志1 王正敏 2 孙珊 2 李华伟2白燕1 1. 重庆医科大学附属儿童医院耳鼻喉科400014 重庆；2.复旦大学附属眼耳鼻喉科医院耳鼻喉科 20013上海 通信作者：白燕，电话 ， Email:baiyan1995@ 【摘要】：目的：研究新霉素致损小鼠的听觉上皮后核修饰基因Mitochondrial transcription factor B1（TFB1M）与听觉上皮线粒体膜电位、毛细胞凋亡的相关性。方法：采用新生ICR小鼠听觉上皮体外无血清培养，建立新霉素致损模型后，随机分为正常组和新霉素致损组，在不同时间点：致损4h，12h，24h，洗脱后2d，检测核修饰基因TFB1M在听觉上皮细胞核和线粒体中的表达，线粒体膜电位（MMP）的变化以及毛细胞的凋亡。结果：新霉素致损组，在致损4 h后，尽管听觉上皮中圈外毛细胞形态完整，但是已经出现线粒体膜电位的改变；新霉素致损12 h，耳蜗听觉上皮线粒体以及细胞核内的TFB1M表达上调，于此同时听觉上皮出现毛细胞从底圈到中圈逐渐缺失，底圈、中圈大量外毛细胞缺失，未缺失处毛细胞出现膜电位明显降低；新霉素致损24 h中圈、底圈毛细胞基本消失，未检测到线粒体膜电位；洗脱后2天，致损组较对照正常组TFB1M表达降低，基底膜毛细胞未见进一步缺失。而正常组在新霉素致损相对应的时间点上，离体培养的听觉上皮的毛细胞形态完整，排列整齐，膜电位无改变。结论： TFB1M蛋白质水平的表达上调可能导致线粒体功能异常和毛细胞凋亡，可能对新霉素类药物导致的药物性耳聋的防治具有潜在重要的意义 【关键词】：TFB1M;线粒体;毛细胞;凋亡 基金项目：重庆市科委基金项目资助（CSTC 2010BB537） 【中文图书分类法分类】R76 【文献标识码】A 【作者投稿日期】2014-08-13 Study of the relationship between nuclear gene TFB1M, auditory epithelial mitochondria and hair cell apoptosis Yang Dazhi1, Wang Zhengmin2, Sun Shan2, Li Huawei2, Bai Yan1. 1.Department of otolaryngology, Childrens Hospital, Chongqing Medical University, Chongqing 400014, China; 2. Department of otolaryngology, Eye ENT Hospital, Fudan university, Shanghai, 20013, China 【Abstract】Objective：In order to observe the relationship between nuclear gene TFB1M, auditory epithelial mitochondria, and hair cell apoptosis in neomycin-induced hair cell death in the mouse in vitro. Methods：First, we establish practical method of auditory epithelia culture model in vitro for P1～P2 mice. Second we detect the mitochondrial membrane potential of auditory epithelia. Then we detect the expression of nuclear gene TFB1M and apoptosis related gene in auditory epithelia of mice. Results：During the first 4h in culture with neomycin treatment, we found the morphogenesis of hair cell in middle turn were aligned, the mitochondrial membrane potential was intact in the majority of the cells. Except few hair cell was indicated as by a slight decrease their membrane potential. Dur