顺铂对离体C57小鼠耳蜗毛细胞的损害作用

Cisplatin-Induced Damage of C57 Mice Hair Cells in Cochlear Organizational Cultures

曾珊;于栋祯;陈正侬;陈斌;

1:上海交通大学附属第六人民医院耳鼻咽喉科

2:上海交通大学耳鼻咽喉科研究所

3:上海交通大学眩晕疾病诊治中心

摘要
目的探讨离体培养条件下顺铂致C57小鼠耳蜗毛细胞损伤的特点。方法出生后3~4天的C57小鼠耳蜗基底膜离体培养6~8小时后,加入不同浓度的顺铂(0、10、50、100、400、1000μmol/L)无血清培养液继续培养48小时,其中0μmol/L为正常对照组,10~1000μmol/L组为实验组,0~100μmol/L4组,每组各7只耳蜗,400和1000μmol/L两组各6只耳蜗。采用TRITC-Phalloidin和DAPI染色,荧光显微镜下观察耳蜗毛细胞生长情况并拍照计数、绘制毛细胞缺失图,运用SARS8.0软件进行直线回归分析比较各组的毛细胞缺失率。结果对照组耳蜗基底膜毛细胞在离体培养条件下生长良好,偶见缺失。实验组中顺铂引起毛细胞的缺失程度从底回至顶回大致均匀。随着顺铂浓度从10μmol/L增加到100μmol/L,毛细胞的缺失率从14.5%上升到78.4%;顺铂浓度升高至400μmol/L及1000μmol/L时,毛细胞缺失率下降至48.8%和8.77%。细胞缺失区域可见DAPI着色的浓缩核。结论离体培养条件下,顺铂对耳蜗毛细胞的损伤在一定浓度范围内呈剂量依赖性,超过一定剂量后顺铂所致毛细胞损伤又逐渐减少;内外毛细胞的损伤从底回至顶回呈现均一性。
关键词
听毛细胞;耳蜗;器官培养;顺铂
基金项目(Foundation):
作者
曾珊;于栋祯;陈正侬;陈斌;
参考文献

1Rybak LP,Whitworth CA,Mukherjea D,et al.Mechanisms of cisplatin-induced ototoxicity and prevention[J].Hear Res,2007,226:157.

2van Ruijven MW,de Groot JC,Smoorenburg GF.Ti me se-quence of degeneration patternin the guinea pig cochlea during cisplatin administration.A quantitative histological study[J].Hear Res,2004,197:44.

3Cheng AG,Cunningham LL,Rubel EW.Mechanisms of hair cell death and protection[J].Curr Opin Otolaryngol Head Neck Surg,2005,13:343.

4Lutsenko S,Petris MJ.Function and regulation of the mam-malian copper-transporting ATPases:insights from bioche-mical and cell biological approaches[J].J Membr Biol,2003,191:1.

5Safaei R.Role of copper transporters in the uptake and efflux of platinumcontaining drugs[J].Cancer Lett,2006,234:34.

6Holzer AK,Katano K,Klomp LW,et al.Cisplatin rapidly down-regulates its owninflux transporter hCTR1in cultured human ovarian carcinoma cells[J].Clin Cancer Res,2004,10:6744.

7van Ruijven MW,de Groot JC,Klis SF,et al.The cochlear targets of cisplatin:an electrophysiological and morphological ti me-sequence study[J].Hear Res,2005,205:241.

8Alam SA,Ikeda K,Oshi ma T,et al.Cisplatin-induced a-poptotic cell deathin Mongolian gerbil cochlea[J].Hear Res,2000,141:28.

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曾珊于栋祯陈正侬陈斌