摘要目的  通过微量联合皮下注射硫酸卡那霉素与腹腔注射速尿的实验方法，建立并鉴定小鼠感音神经性耳聋动物模型，为课题组后续耳聋小鼠药物性干预研究提供模型基础。

方法  1、用硫酸卡那霉素与速尿联合用药建立感音性耳聋动物模型：60只健康CBA/caj雄性小鼠随机平均分成6 组：正常组（空白对照）、给硫酸卡那霉素与速尿联合用药后1d组、用药后3d组、用药后7d组、用药后14d组、用药后28d组，药物模型组皮下注射硫酸卡那霉素和腹腔注射呋塞米给药后仍正常饲养，10只小鼠作为对照组不予任何处理；2、运用听性脑干反应测试（auditory brainstem response，ABR）和HE染色组织形态学进行感音性耳聋模型的鉴定：小鼠用药前和用药后1d、3d、7d、14d、28d使用美国TDT进行听性脑干反应（auditory brainstem response ABR）检测。各组小鼠于在进行 ABR 检测后，通过耳蜗基底膜HE染色铺片显微镜下观察小鼠耳蜗基底膜病理切片中毛细胞缺失和排列，并摄片记录。88131

结果  1）ABR结果显示，致聋前，对照组和药物致聋组的ABR阈值<40dB,致聋后，对照组的ABR阈值<40dB，药物致聋组的ABR阈值>70dB,药物致聋组联合用药后对小鼠可引起听力明显下降，造成小鼠永久性听阈位移，听阈提高超过40dB。且速尿和硫酸卡那霉素致聋前后对照组ABR阈值差异无统计学意义(P>0。05)，致聋前后对照组与药物致聋组之间小鼠的ABR 阈值差异有统计学意义(P<0。01)；2）耳蜗基底膜HE染色铺片观察显示，随着硫酸卡那霉素损伤时间的延长，耳蜗外毛细胞的排列越来越紊乱 ，建模后一天外毛细胞出现轻微的紊乱，建模后28天小鼠耳蜗毛细胞已看不出原来的排列方式。源-于,优W尔Y论L文.网wwW.youeRw.com 原文+QQ75201,8766

结论  速尿和硫酸卡那霉素联合用药后可导致小鼠永久性听阈位移，可使耳蜗毛细胞损细胞排列紊乱，且给药时间的长短对毛细胞的损伤程度有一定的影响，通过联合注射速尿和硫酸卡那霉素可建立理想的小鼠药物耳聋动物模型。

毕业论文关键词：毛细胞； 小鼠动物模型； 感音神经性耳聋

Abstract Objective this study aimed to establish a model of sensorineural deafness in neonatal mouse by co -administration of the loop diuretic furosemide and kanamycin to inner skin and enterrocolia using a nanlolite micro-injection system。explore the effective method of establishing animal models of sensorineural deafness。 　

Methods 60mice were randomly pided into 6 groups。 10 mice served as the control group without receiving anthing,The mice of rest group received an co-administration of the loop diuretic furosemide and kanamycin to inner skin and enterrocolia ,auditory brainstem response（ABR）were recorded to monitor the animal's hearing at the 1day,3days,7days,14days,28days after the drug administration．And detect the pathological damage process of hair cell with the surface preparation techniques of the cochlear basilar membrane after ABR detection。 

Results  control group and drug group deafness ABR threshold < 40 dB。

deafness, tHE control of ABR threshold < 40 dB, drug deafness group of ABR thresholds > 80 dB, drug deafness group after combination in mice can cause hearing declined obviously, causing permanent displacement of auditory threshold in mice, auditory threshold increased more than 40 dB。 Compared with the control group, ABR threshold of t 5 experi-mental groups were significantly higher, and can cause tHE damage of hair cells after co-administration of tHE loop diuretic furosemide and kanamycin to inner skin and enterrocolia 。

Conclusion Furosemide and kanamycin sulfate can lead to permanent displacement of auditory threshold in mice after combination, can make the disordered arrangement of cochlear hair cells loss, and the dosage of the length of time that the damage degree of hair cells have a certain influence。