C57BL／6J小鼠耳蜗质膜钙ATP酶异构体2的年龄相关性表达  

Age-related expression of plasma membrane Ca2＋ -ATPase isoform 2 in the cochleas of C57BL/6J mice

作　　者：高燕[1] 陶雁玲[1] 褚汉启[1] 陈金[1] 陈请国[1] 周良强[1] 刘云[1] 余洋[1] 崔永华[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院耳鼻咽喉头颈外科,武汉430030

出　　处：《中华耳鼻咽喉头颈外科杂志》2015年第11期934-938,共5页Chinese Journal of Otorhinolaryngology Head and Neck Surgery

基　　金：国家自然科学基金（81271078,81070787,81300827）;同济医院科研基金（2201101861）

摘　　要：目的研究质膜钙ATP酶异构体2（plasmamembraneCa2＋-ATPaseisoform2，PMCA2）在C57BL／6J小鼠耳蜗组织中的分布及年龄相关性表达，探讨其与年龄相关性听力损失的关系。方法运用免疫荧光组织化学的方法检测PMCA2蛋白在不同鼠龄（4、14、22、45周）C57BL／6J小鼠耳蜗中的分布和表达变化；采用实时定量多聚酶链反应（Real-timePCR）检测PMCA2mRNA在不同鼠龄（4、14、22、45周）小鼠耳蜗中的表达。采用SPSS17．0软件进行统计学分析。结果免疫荧光的结果显示不同鼠龄C57BL／6J小鼠的耳蜗内外毛细胞、血管纹和螺旋神经节细胞均可见PMCA2蛋白表达，螺旋韧带亦可见有少量PMCA2蛋白表达。随着鼠龄的增长，耳蜗毛细胞出现缺失，螺旋神经节细胞数目逐渐减少，PMCA2蛋白的表达也随着鼠龄的增加而逐渐减少。Real—timePCR结果揭示：随着鼠龄的增长，PMCA2mRNA表达水平逐渐下降，14周龄的表达水平与4周龄相比，差异有统计学意义（P〈0．05）；22周龄表达水平与14周龄相比，差异有统计学意义（P〈0．05）；45周龄表达水平与14周龄相比，差异有统计学意义（P〈0．01）。结论PMCA2蛋白主要分布于C57BL／6J小鼠耳蜗内外毛细胞、血管纹和螺旋神经节细胞，随着鼠龄的增长，PMCA2蛋白和mRNA的表达量逐渐减少。PMCA2的表达下降或功能缺失可能与C57BL／6J小鼠内耳形态的年龄相关性改变以及听力损失有一定关系。Objective To investigate the location and distribution of plasma membrane Ca2＋- ATPase isoform 2 （PMCA2） in the cochleas of C57BL/6J mice at various ages （4w, 14w, 22w, 45w） , and to reveal the relationship of PMCA2 and age-related hearing loss （AHL）. Methods The distribution of PMCA2 in the cochleas of C57BL/6J mice was detected by immunohistochemistry at various ages （4w, 14w, 22w, 45w）. Real-time polymerase chain reaction （Rt-PCR） was used to detect the level of PMCA2 mRNA in the cochleas of C57BL/6J mice at the ages of 4, 14, 22 and 45 weeks old respectively. Using SPSS17.0 software for statistical analysis. Results PMCA2 was mainly located in the hear cells, stria vascularis, and spiral ganglion cells. Faint labeling of PMCA2 was also observed in spiral ligament. Hair cells missed and the number of spiral ganglion cells reduced with age. Expression of PMCA2 in the cochleas of C57BL/6J mice also showed age-related decreasing. The results of Rt-PCR demonstrated the expression of mRNA of gene （ Atp2b2 ） at 14 weeks age was significantly less than 4 week-old mice eochlears （ P 〈 0. 05 ）. The expression of mRNA of gene （ Atp2b2 ） at 22 weeks age was significantly less than 14 week-old mice cochlears（P 〈0. 05）. The expression of mRNA of gene （Atp2b2） at 45 weeks age was significantly less than 14 week-old mice cochlears （ P 〈 0. 01 ）. Conclusions PMCA2 is mainly located in the hear cells,stria vascularis, and spiral ganglion cells. Faint labeling of PMCA2 is also observed in spiral 1 igament. The expression of PMCA2 demonstrates an age-related decrease with age. The mRNA expression level of PMCA2 gene （ Atp2b2 ） in the cochleas of C57BL/6J mice displayed an age-related decrease. PMCA2 transporters may play a critical role in maintaining the normal morphology of the inner ear and it may be related to AHL.

关 键 词：耳蜗 质膜钙转运ATP酶类 老年性聋 小鼠 近交C57BL 

分 类 号：R764.43[医药卫生—耳鼻咽喉科]