硫酸卡那霉素联合呋塞米快速致聋大鼠模型的研究

Acute Deafness Induced by Co-administration of Kanamycin Sulfate and Furosemide in a Rat Animal Model

闫辉;邢蔚;宋勇莉;王人凤;陈俊;邱建华;

1:第四军医大学西京医院耳鼻咽喉头颈外科

2:新疆生产建设兵团医院(石河子大学医学院第二附属医院)耳鼻咽喉科

3:兰州军区临潼疗养院第二疗养区耳鼻咽喉科

摘要
目的观察硫酸卡那霉素(kanamycin sulfate,KM)与呋塞米(furosemide,Fur)联合用药建立药物性快速致聋大鼠动物模型的效果。方法 32只SD大鼠随机分为A组(KM+Fur后1天组)、B组(KM+Fur后7天组)、C组(KM+Fur后14天组)、D组(对照组),每组8只动物。A、B、C三组分别经一次性腹腔注射KM 500mg/kg,30分钟后再经腹腔注射Fur 0.2ml/kg,空白对照组按体重给予相同剂量生理盐水。各组在相应的时间点完成听性脑干反应(auditory brainstem response,ABR)检测当天,分别行耳蜗基底膜铺片及扫描电镜观察耳蜗毛细胞损伤情况,免疫荧光染色观察螺旋神经节细胞亡情况。结果 A、B、C组各频率ABR阈值较正常对照组明显升高(P<0.05);而A、B、C组各频率ABR反应阈差异无统计学意义(P>0.05)。A、B、C组耳蜗底回外毛细胞明显缺失,纤毛排列紊乱,螺旋神经节细胞的半胱氨酸天冬氨酸蛋白酶-3(cleaved caspase-3)的表达量较正常对照组均明显增加(P<0.05)。结论硫酸卡那霉素和呋塞米联合单次腹腔用药24小时后大鼠ABR反应阈明显增高,并可见耳蜗底回毛细胞的明显缺失,KM+Fur联合应用是简单、快速而有效的大鼠耳聋动物模型建模方法。
关键词
硫酸卡那霉素;呋塞米;大鼠;耳聋;动物模型
基金项目(Foundation):
国家自然科学基金青年项目(81100713);国家自然科学基金海外重大合作项目(81120108008);; 国家“973”项目课题(2011CB504505)资助
作者
闫辉;邢蔚;宋勇莉;王人凤;陈俊;邱建华;
参考文献

1 Zhou Y,Ding D,Kraus KS,et al.Functional and structural changes in the chinchilla cochlea and vestibular system following round window application of carboplatin[J].Audiol Med,2009,7:189.

2 He J,Yin S,Wang J,et al.Effectiveness of different approaches for establishing cisplatin-induced cochlear lesions in mice[J].Acta Otolaryngol,2009,129:1 359.

3 丁大连,蒋涛,丌卫东,等.内耳科学[M].北京:中国科学技术出版社,2010.222~223.

4 West BA,Brummett RE,Himes DL.Interaction of kanamycin and ethacrynic acid.Severe cochlear damage in guinea pigs[J].Arch Otolaryngo1,1973,98:32.

5 Nourski KV,Miller CA,Hu N,et al.Co-administration of kanamycin and ethacrynie acid as a deafening method for acute animal experiments[J].Hear Res,2004,187:131.

6 Astbury PJ.Read NG.Kanamycin induced ototoxicity in the laboratory rat:a comparative morphological and audiometric study[J].Arch Toxicol,1982,50:267.

7 Murillo-Cuesta S,Garcia-Alcfintara F,Vacas E,et al.Direct drug application to the round window:a comparative study of ototoxicity in rats[J].Otolaryngol Head Neck Surg,2009,141:584.

8 Wu WJ,Sha SH,McLaren JD,et al.Aminoglycoside ototoxicity in adult CBA,C57BL and BALB mice and the SpragueDawley rat[J].Hear Res,2001,158:165.

9 张贤芬,杨仕民,胡吟燕,等.联合应用卡那霉素和速尿的豚鼠耳蜗毒性实验观察[J].中华耳科学杂志,2008,6:166.

10 Gibbs RA,Weinstock GM,Metzker ML.Genome sequence of the brown Norway rat yields insights into mammalian evolution[J].Nature,2004,428:475.

11 Yang JH,Liu ZH.Apoptosis of cochlear cells and hearing threshold in guinea pigs with kanamucin-induced chronic intoxication[J].Chinese Journal of Clinical Rehabilitation,2005,9:30.

12 杨俊慧,刘兆华.卡那霉素耳慢性中毒后豚鼠耳蜗细胞凋亡的观察[J].中国耳鼻咽喉科杂志,2004,4:91.

13 He J,Yin S,Wang J,et al.Effectiveness of different approaches for establishing cisplatin-induced cochlear lesions in mice[J].Acta Otolaryngol,2009,12:1 359.

14 Geal-Dor M,Freeman S,Li G,et al.Development of hearing in neonatal rats:air and bone conducted ABR thresholds[J].Hear Res,1993,69:236.

15 邬莎,王爱梅.Caspase家族与内耳细胞凋亡[J].国际耳鼻咽喉头颈外科杂志,2007,6:317.

16 Jad W,Cristina P,Fiona LS,et al.A constitutively active and uninhibitable caspase-3zymogen efficiently induces apoptosis[J].Biochem J,2009,424:335.

17 Suparna M,Dragos P,Alexandru A,et al.Caspase-3activation is a critical determinant of genotoxic stress-induced apoptosis[J].Methods in Molecular Biology,2008,414:13.

18 Shepherd JD,Bear MF.New views of Arc,a master regulator of synaptic plasticiticy[J].Nat Neurosci,2011,14:279.