庆大霉素对内毛细胞带状突触CaV1.3钙离子通道蛋白表达的影响  

Gentamicin on inner hair cells ribbon synapases CaV1.3 calcium ion channel protein expression

作　　者：孙建华[1] 王雪峰[2] 柳柯[3] 

机构地区：[1]锦州市中心医院耳鼻咽喉头颈外科,辽宁锦州121000 [2]辽宁医学院附属第一医院耳鼻咽喉头颈外科 [3]中国人民解放军总医院耳鼻咽喉头颈外科

出　　处：《临床耳鼻咽喉头颈外科杂志》2014年第4期261-264,共4页Journal of Clinical Otorhinolaryngology Head And Neck Surgery

摘　　要：目的:研究庆大霉素对C57BL/6J小鼠听力及耳蜗内毛细胞带状突触CaV1.3钙通道蛋白数量的影响,探讨听力损失与其剂量相关性变化之间的关系及意义。方法:采用固定剂量[100mg/(kg·d)]的氨基苷类抗生素(庆大霉素)对C57BL/6J小鼠每日进行腹腔注射造模,分别在注射后的第7天、第14天、第28天(注射14d停药14d)应用ABR分别测得小鼠的听力。免疫荧光方法观察耳蜗基膜内毛细胞带状突触CaV1.3钙通道蛋白的分布和表达(其中0天为空白对照组),内毛细胞突触前后膜分别采用CtbP2和CaV1.3进行免疫荧光标记。结果:随着注射药物天数的增长ABR反应阈值明显提高,所有小鼠内毛细胞数量、形态均无明显变化,但内毛细胞带状突触CaV1.3钙离子通道蛋白量的表达存在显著差异(P<0.01)。CaV1.3钙离子通道蛋白在庆大霉素腹腔注射CaV1.3/CtBP2表达数量在第7天时略有增加,在第14天时其表达数量明显减少,第28天与第14天比较CaV1.3/CtBP2表达数量有所增加,但低于正常。结论:在氨基苷类抗生素毒性环境中耳蜗内毛细胞CaV1.3蛋白表现为先增加后减少再增加的趋势,提示内毛细胞带状突触CaV1.3蛋白数量的增加可能存在对这类药物毒性的代偿作用,而随着药物毒性作用时间的增加,这种失代偿作用表现为听力下降,可能是损伤听力的机制之一。Objective：To learn the influence the gentamycin on C57BL/6J mice hear and cochlear hair cell rib-bon synapses CaV1.3 calcium protein amount. To explore the relationship between hear loss and its dosage correla- tion change and significance. Method： The fixed amino glucoside to C57BL/6] mice was used to make abdominal cavity injection mold every day. The auditory brain-stem response ABR was used to measure the hear of mice in 7th, 14th, 28th after the injection. Immunofluorescence method was used to observe cochlear basement membrance of hair ribbon synapse CaV1.3 calclum channel proteins in the distribution and expression. Inner hair cells synaptic membrane was immune fluorescent tags with CtbP2 and CaV1.3. Result： With the growth of the injected drugs, ABR threshold increased,but all the hair cells and shape had no obvious change. However the amount of hair rib- bon synapse CaV1.3 calcium ion channel proteins in the expression had significant differences（P^0.01 ）. CaV1.3 caicium ion channel proteins increased slightly lower than normal at 7th day, significantly decreased at 14th day, had increased,increased quantity compare with 14th day,but at 28th day after intraperitoneal injection of gentami- cin. Conclusion：The increasing,decreasing and increasing trend of cochlear hair cells CaV1.3 proteins in the envi- ronment of amino glucoside drug toxicity showed that the increase of hair ribbon synapse CaV1.3 proteins may have a compensatory effect on the drug toxicity. With the increase of the drug toxicity effect, this kind of decom- pensated function could be the listening decline,which may be one of the mechanism of damage to hearing.

关 键 词：庆大霉素 耳蜗 内毛细胞 带状突触 听性脑干反应 CaVl 3 

分 类 号：R764[医药卫生—耳鼻咽喉科]