卡那霉素对体外培养小鼠耳蜗毛细胞的损伤作用  

Impairment Effect of Kanamycin on Mouse Cochlear Hair Cells in Vitro

作　　者：许涛[1] 王爱梅[2] 刘双月[1] 牟华[2] 

机构地区：[1]辽宁医学院研究生院 [2]辽宁医学院生理学教研室,辽宁锦州121001

出　　处：《江西医学院学报》2009年第11期18-20,F0004,共4页Acta Academiae Medicinae Jiangxi

基　　金：辽宁省教育厅科学研究计划资助项目(202173374);辽宁省博士科研启动基金资助项目(20031086)

摘　　要：目的建立新生昆明小鼠耳蜗Corti器体外培养模型,研究卡那霉素(kanamycin,KM)对体外培养小鼠耳蜗毛细胞的损伤作用。方法分离生后3d的小鼠耳蜗基底膜,平铺在胶原凝胶表面,然后在含有1%牛血清白蛋白的DMEM/F12培养基中进行培养。采用异硫氰酸四甲基罗丹明标记的鬼笔环肽荧光染色方法观察不同浓度(0.05、0.1、0.2、0.3mmol/L)KM对小鼠耳蜗毛细胞的影响,耳蜗毛细胞计数检测毛细胞缺失情况。结果不同浓度KM均可使小鼠耳蜗外毛细胞出现纤毛倒伏、排列紊乱等形态变化。KM浓度为0.05mmol/L时,耳蜗外毛细胞缺失率明显增大,与对照组比较有显著性差异(P<0.05),而耳蜗内毛细胞缺失率无明显变化(P>0.05)。随着KM浓度的逐渐增高(0.1～0.3mmol/L),耳蜗内、外毛细胞缺失率亦显著增大,呈现明显的量效关系(P<0.05或P<0.01)。结论KM可对体外培养的小鼠耳蜗毛细胞产生损伤作用,内耳器官培养是研究药物耳毒性的有效方法。Objective To establish an organ culture model of Corti to investigate the impairment effect of kanamycin（KM）on mouse cochlear hair cells.Methods The cochlear basilar membrane from 3-day-old mouse were isolated and positioned on the surface of collagen gel and cultured in DMEM/F12 medium containing 1% bovine serum albumin.Tetramethylrhodamine isothiocyanate-labeled phalloidine staining was used to observe the effects of different concentration of KM（0.05,0.1,0.2,0.3 mmol/L）on mouse cochlear hair cells.The loss of hair cells was detected by cell counts.Results The cilia of cochlear outer hair cells were lodged and arranged in disorder caused by different concentration of KM.When KM concentration of was 0.05 mmol/L,the percent of missing outer hair cell was significantly increased as compared with control group（P〈0.05）,while the percent of missing inner hair cell was no significant changed（P〉0.05）.With KM concentration was gradually increased（0.10.3 mmol/L）,the percents of missing inner and outer hair cell were significantly increased,showing a clear dose-esponse relationship（P〈0.05 or P〈0.01）.Conclusion KM could impair hair cells in mouse cochlear culture,and organ culture of inner ear is a effective method for studying drug ototoxicity.

关 键 词：卡那霉素 耳蜗 毛细胞 体外培养 动物 实验 小鼠 

分 类 号：R764.43[医药卫生—耳鼻咽喉科]