中文摘要：

目的研究有机氧化剂叔丁基过氧化氢（t-BHP）体外模拟噪声对耳蜗毛细胞的氧化损伤。方法用t-BHP染毒，设置从30-4000μmol／L8个染毒浓度对耳蜗毛细胞（HEI-Ocl）细胞染毒12h，绘制100μmol／L浓度组染毒时间（1-95h）与耳蜗毛细胞存活率曲线；采用台盼蓝染色法检测细胞存活率，噻唑蓝（M1Tr）试验检测细胞增殖能力的改变，2＇-7’-二氯荧光黄双乙酯（DCFH-DA）探针法检测胞内活性氧（ROS）水平。结果不同浓度的t-BHP对耳蜗毛细胞染毒12h后，100μmol／L以上浓度组细胞存活率开始出现有统计学意义的下降，其中200．2000Ixmol／L浓度组细胞存活率呈直线下降。100μmol／L浓度组t-BHP染毒时间一细胞存活率曲线较为平缓。MTF试验提示，30μmol／L的t-BHP染毒可以促进耳蜗毛细胞增殖．200μmol／L以上各浓度组则抑制细胞增殖。DCFH—DA探针法显示，无染毒细胞镜下无荧光（-）．阳性对照为强荧光（＋），30和50μmol／L浓度组则有弱荧光（--＋），100μmol／L染毒组为强明亮荧光（＋＋）．200-1000μmolL各组荧光均较强，但随着存活率的迅速降低视野下细胞稀疏。结论100μmol／L的t-BHP对HEI．OCl细胞染毒12h可以在不影响细胞外形、增殖能力的前提下提高耳蜗毛细胞内ROS水平，模拟噪声引起的氧化应激。

英文摘要：

Objective To establish the oxidative damage model of cochlea hair ceils using organic oxidant t-BHP in vitro. Methods HEI-OCl cells were exposed to t-BHP at 8 doses （30-4000 μmol/L） for 12 h. Trypan blue test was used to detected the cellular viability and MTY assay was utilized to measured the cellular proliferation. The intracellular ROS levels were determined by 2, 7-dichlorodihydrofluorescein diacetate （DCFH-DA）. Results The survival rates of HEI-OC1 cells started decrease significantly at the dose of 100μmol/L t-BHP, the peak of decreased survival rates appeared at the doses of 200-800μmol/L. The results of MTF assay demonstrated that 30μmol/L t-BHP could promote cellular proliferation ability, when t-BHP concentrations were higher than 200 μmol/L, the cellular proliferation ability was inhibited. The results of DCFH-DA assay showed that there was no fluorescence in control group, the strong fluorescence was observed in positive control group, the weak fluorescence was observed in 30-50μmol/L t-BHP groups, the bright fluorescence was observed in 100 μmol/L t-BHP group, still the stronger fluorescence was observed in 200-1000 μmol/L groups, but the cellular number decreased with the doses because of the lower cellular viability. Conclusion The exposure to 100 μmol/L t-BHP for 12 h could simulate the oxidative damage induced by