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

作　　者：曾珊[1] 于栋祯[1] 陈正侬[1] 陈斌[1] 

机构地区：[1]上海交通大学附属第六人民医院耳鼻咽喉科,上海交通大学耳鼻咽喉科研究所,上海交通大学眩晕疾病诊治中心,上海200233

出　　处：《听力学及言语疾病杂志》2010年第4期367-371,共5页Journal of Audiology and Speech Pathology

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。Objective To investigate the characteristic of the cisplatin--induced damage of hair cells in cochlear organotypic cultures. Methods Three to four--day--old （P3-4） C57 mice were used for this study. The cochlear basilar membrane was cultured for 6-8 hours and then treated with different doses of cisplatin（0μmol/L, 10μmol/L, 50μmol/L, 100μmol/L, 400μmol/L, 1000μmol/L,7 cochlears for 0--100μmol/L group, 6 cochlears for the rest dosage, control group used the dose of 0μmol/L）for 48 hours. TRITC- phalloidin and DAPI were used for stain. The entire cochlear hair cells were observed and counted under Nicon fluorescent microscope. Data was analyzed by LS for hair cells loss graph and SAS 8.0 for linear regression analysis. Results Hair cells loss had rarely been seen in the control group of cochlear tissue under the culture condition. How ever, in cisplatin treated groups, the hair cells loss increased from 14. 5% to 78. 4% with the concentration of cisplatin increased in the range of 10μmol/L-100μmol/L, while decreased from 48. 8% down to 8. 77% with the dose getting further higher to 400μmol/L and 1000μmol/L. Apoptosis was confirmed by the DAPI stain in the hair cells loss area. The hair cell loss shows no difference from the apex to the basal turn of the basilar membrane. Conclusion Conclatin--induced hair cell death shows characteristic damage that it is dose--dependent in a certain range of dosage, and it decreases when the cisplatin dose higher than the one caus maximal damage under the condition of cochlear organotypic cuturcs. Apoptosis happens evenly in inner and outer hair cells in all turns of basilar membrane.

关 键 词：听毛细胞 耳蜗 器官培养 顺铂 

分 类 号：R764.5[医药卫生—耳鼻咽喉科]