卡那霉素和2-羟丙基-β-环糊精耳毒性的比较研究

doi:10.6040/j.issn.1673-3770.0.2021.195

山东大学耳鼻喉眼学报 ›› 2022, Vol. 36 ›› Issue (4): 6-11.doi: 10.6040/j.issn.1673-3770.0.2021.195

卡那霉素和2-羟丙基-β-环糊精耳毒性的比较研究

杨琨¹,陈利娟²,何小丹³,刘志奇³,沙素华⁴

1. 武汉大学人民医院耳鼻咽喉头颈外科, 湖北武汉 430060;
2. 华中科技大学附属湖北省妇女儿童医院儿童神经内科, 湖北武汉 430075;
3. 华中科技大学附属湖北省妇女儿童医院耳鼻咽喉科, 湖北武汉 430075;
4. 南卡罗莱纳医科大学病理科, 南卡罗莱纳州查尔斯顿 29425

出版日期:2022-07-20 发布日期:2022-07-11
通讯作者: 刘志奇. E-mail: liuwu188@sina.com
基金资助:
湖北省自然科学基金指导性计划项目(2018CFC839);湖北省卫生健康委员会面上项目(WJ2019M230);湖北省卫生计生委联合基金面上项目(WJ2018H0143);第四批武汉中青年医学骨干人才培养工程(22092007604);湖北省医学拔尖人才项目(1020012203)

Comparative study of ototoxicity between kanamycin and 2-hydroxypropyl-β-cyclodextrin

YANG Kun¹, CHEN Lijuan², HE Xiaodan³, LIU Zhiqi³, SHA Suhua⁴

1. Department of Otorhinolaryngology & Head and Neck Surgery, People's Hospital of Wuhan University, Wuhan 430060, Hubei, China;
2. Department of Neurology, Hubei Women's and Children's Hospital, Huazhong University of Science and Technology, Wuhan 430070, Hubei, China;
3. Department of Otorhinolaryngology, Hubei Women's and Children's Hospital, Huazhong University of Science and Technology, Wuhan 430070, Hubei, China;
4. Department of Pathology and Laboratory Medicine, Medical University of South Carolina, Charleston, SC 29425, USA

Online:2022-07-20 Published:2022-07-11

摘要/Abstract

摘要： 目的初步比较研究卡那霉素和2-羟丙基-β-环糊精(HPβCD)致CBA/J小鼠耳毒性损伤的特点。方法雄性CBA/J小鼠,700 mg/kg卡那霉素连续腹腔注射15 d,HPβCD8 000 mg/kg皮下注射,对照组均为0.9%无菌氯化钠生理盐水。听性脑干反应(ABR)检测听力。耳蜗基底膜铺片和二氨基联苯胺染色法(DAB)毛细胞计数。HPβCD多个时间点听阈的比较和外毛细胞记数的比较使用统计软件GraphPad Prism 13 中重复测量方差分析法(Repeated measures ANOVA)进行数据分析,卡那霉素实验中听阈的比较用GraphPad Prism 13中的双样本t检验(T-Test)。结果生理盐水组8 kHz、16 kHz、32 kHz阈值分别为(21±2.24)dB、(21±2.24)dB、(21±2.24)dB;卡那霉素治疗组阈值分别为(22±2.74)dB(t=0.632 5,P=0.544 7)、(67±4.47)dB(t=20.571 8,P<0.001)、(77±4.47)dB(t=25.044 0, P<0.001)。HPβCD治疗组6 h后,8 kHz、16 kHz、32 kHz反应分别为(29±4.18)dB、(30±4.47)dB、(37±2.74)dB,7h时后分别为(65±3.54)dB、(71±2.24)dB、(80±3.54)dB,8 h后分别为(70±3.54)dB、(78±5.70)dB、(85±5.00)dB,经比较显示差异有统计学意义(F=20.590,P<0.001)。随着HPβCD用药时间的延长,各频率听力损失逐渐加重。卡那霉素治疗组OHC损失于顶转至基底转2.0 mm处为(9.17±6.03)%,2.5 mm处为(89.76±3.12)%, 4.0 mm处为100%,经比较显示差异有统计学意义(F=7.840,P=0.000 3);IHC损失分别为(0.71±0.00)%、(6.79±6.01)%、(20.71±6.00)%,经比较显示2.5 mm和4.0 mm处差异有统计学意义(F=3.549,P<0.05)。HPβCD治疗6 h后耳蜗基底膜外毛细胞可见明显的损伤,外毛细胞的损伤在基底膜顶转,中转和底转都可出现,7、8 h后耳蜗内外毛细胞几乎完全脱落,8 h以内未见明显的内毛细胞损伤。结论卡那霉素和HPβCD均致小鼠听力下降、毛细胞损伤。卡那霉素耳毒性损伤从高频开始,HPβCD耳毒性听力损伤在早期就波及全频,外毛细胞损伤先于内毛细胞损伤,损伤程度都具有时间和浓度依赖性。

关键词: 药物性聋, 内毛细胞, 外毛细胞, 2-羟丙基-β-环糊精, 卡那霉素, 耳毒性

Abstract: Objective This study aimed to investigate the effects of ototoxic injury induced by kanamycin and 2-hydroxypropyl-β-cyclodextrin(HPβCD)in CBA/J mice. Methods Male CBA/J mice were intraperitoneally administered kanamycin for 15 days at the optimal dosage of 700 mg/kg and were subcutaneously administered HPβCD at the optimal dosage of 8 000 mg/kg. The control group was administered 0.9 % sterile sodium chloride. Auditory brainstem response tests were performed. Cochlear preparations and diaminobenzidine staining were used for hair cell observation. Data were analyzed using the statistical software GraphPad Prism 13 for Windows. Results In the control group, at 8 kHz, 16 kHz, and 32 kHz, the thresholds were 21±2.24 dB, 21±2.24 dB, and 21±2.24 dB, respectively. After kanamycin treatment, the thresholds were 22±2.74 dB(t=0.632 5, P=0.544 7), 67± 4.47 dB(t=20.571 8, P<0.001), and 77±4.47 dB(t=25.044 0, P<0.001), respectively. After 6 hours, the thresholds in the HPβCD group were 29±4.18 dB, 30±4.47 dB, and 37±2.74 dB, respectively. After 7 hours, the thresholds were 65±3.54 dB, 71±2.24 dB, and 80±3.54 dB, and after 8 hours, the thresholds were 70±3.54 dB, 78±5.70 dB, and 85±5.00 dB, respectively. Comparison of the data revealed significant differences(F=20.590, P<0.001). Prolonging HPβCD treatment resulted in hearing loss being gradually aggravated at all frequencies. In the kanamycin group, outer hair cell(OHC)loss from the top to the base was significantly different(F=7.840, P=0.000 3), with 9.17±6.03% at 2.0 mm, 89.76±3.12% at 2.5 mm, and 100% at 4.0 mm. Inner hair cell(IHC)loss was also significantly different(F=3.549, P<0.05), with values of 0.71±0.00%, 6.79±6.01%, and 20.71±6.00% being recorded. Six hours of HPβCD treatment resulted in obvious damage to OHCs of the cochlea. Damage occurred at the apex, the middle, and the base of the basement membrane. After 7 to 8 hours, the IHCs and OHCs had practically disappeared. After 8 hours, no IHCs were found. Conclusion Kanamycin and HPβCD induced hearing loss and hair cell damage in mice. OHC damage occurred earlier than the IHC damage, with the degree of damage being dependent on the time and concentration of treatment.The difference is that Kanamycin damaged hearing at the onset of high frequencies, while HPβCD starts from full frequency.

Key words: Drug-induced hearing loss, DIHL, Inner hair cells, Outer hair cells, 2-hydroxypropyl-β-cyclodextrin, Kanamycin, Ototoxity

中图分类号:

R764.4

杨琨, 陈利娟, 何小丹, 刘志奇, 沙素华. 卡那霉素和2-羟丙基-β-环糊精耳毒性的比较研究[J]. 山东大学耳鼻喉眼学报, 2022, 36(4): 6-11.

YANG Kun, CHEN Lijuan, HE Xiaodan, LIU Zhiqi, SHA Suhua. Comparative study of ototoxicity between kanamycin and 2-hydroxypropyl-β-cyclodextrin[J]. Journal of Otolaryngology and Ophthalmology of Shandong University, 2022, 36(4): 6-11.

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卡那霉素和2-羟丙基-β-环糊精耳毒性的比较研究

Comparative study of ototoxicity between kanamycin and 2-hydroxypropyl-β-cyclodextrin

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