顺铂致聋后大鼠耳蜗螺旋神经元NeuroD的表达  

Expression of NeuroD in spiral ganglion neurons following cisplatin-induced ototoxicity in rats

作　　者：王娟[1] 王亚菲[1] 蒋兴旺[1] 米文娟[1] 邱建华[1] 

机构地区：[1]第四军医大学西京医院全军耳鼻咽喉头颈外科中心,西安710032

出　　处：《中华耳科学杂志》2012年第2期241-245,共5页Chinese Journal of Otology

基　　金：国家973项目(2011CB504505);国家自然科学基金重点项目(30930098);国家自然科学基金海外重大合作项目(81120108008)

摘　　要：目的检测神经分化因子NeuroD在大鼠耳蜗螺旋神经元顺铂损伤后的表达变化。方法通过免疫组织化学及Real-TimePCR技术,观察耳蜗螺旋神经元损伤1d、3d、5d后NeuroD的表达变化。正常成年SD大鼠32只随机分为对照组(生理盐水5ml/kg,1次/d,腹腔注射,连续5d),用药1d组(顺铂5mg/kg,腹腔注射),用药3d组(顺铂5mg/kg,1次/d,腹腔注射,连续3d),用药5d组(顺铂5mg/kg,1次/d,腹腔注射,连续5d),每组8只,建立顺铂耳毒性模型。采用Real-TimePCR、免疫组织化学染色检测不同时间螺旋神经元NeuroD的mRNA及蛋白表达变化。结果成功建立顺铂耳毒性大鼠模型,随着用药时间的延长,神经分化因子NeuroD在耳蜗螺旋神经元中呈动态变化。NeuroD的mRNA和蛋白表达在用药1d及3d组分别为2.17±0.39、1.15±0.20及7.02±0.69、2.42±0.40,与对照组及用药5d组比较具有统计学意义(P值<0.01)。结论 NeuroD在用药1d后开始增加,3d后达到高峰,5d后下降;在用药早期有一过性表达增强,后期表达下降同时听力损失明显。表明NeuroD可能参与顺铂损伤螺旋神经元后的修复过程。Objective To investigate the effects of neurogenic differentiation （NeuroD） on spiral ganglion cells neu- rons（SGNs） in cisplatin-indueed ototoxieity. Methods Thirty two adult Sprague-Dawley（SD）rats were divided randomly into 4 groups （n--8 in each group） to receive intraperitoneal injection of cisplatin （Smg/kg, qd） for 1, 3 or 5 days, or equivalent vol- ume of saline （control group）. The mRNA level of NeuroD expression in cochlear tissues was examined by Real-Time PCR. The expressing pattern of NeuroD in damaged cochlea was studied by immunochemistry using antibodies against NeuroD pro- tein. Results This was a successful model of cisplatin ototoxicity, showing dynamic changes in expression of NeuroD in SGNs along with eisplatin administration. At day 1 and day 3 of eisplatin treatment, NeuroD mRNA levels were 2.17＋0.39 and 1.15 ~0.20, and NeuroD protein levels were 7.02~0.69 and 2.42~0.40, respectively, higher the control and 5 day treatment groups （P〈0.01）. Conclusions Expression of NeuroD seems to increase and peak at days 1-3 after cisplatin treatment, and decrease at day 5. This temporary increase at early stage followed by a later decrease （seemingly parallel to increasing hearing damage） may represent NeuroD＇ s role in the repair of SGNs following cisplatin-induced damage.

关 键 词：NEUROD 大鼠 耳蜗 螺旋神经元 

分 类 号：R322.923[医药卫生—人体解剖和组织胚胎学] R978.12[医药卫生—基础医学]