氨基糖苷类抗生素在小鼠耳蜗毛细胞中的分布特征研究  

Distribution Characteristics of Aminoglycoside Antibiotics in Mouse Cochlear Hair Cells

作　　者：吴俪媛 郭瑞 柳柯[2] 冀飞[3] 乔月华 时晰 WU Liyuan;GUO Rui;LIU Ke;JI Fei;QIAO Yuehua;SHI Xi(Institute of Audiology and Balance Science,Xuzhou Medical University,Xuzhou 221004,China;Department of Otolaryngology,Head and Neck Surgery,Beijing Friendship Hospital,Capital Medical University,Beijing 100000,China;Department of Otolaryngology Head and Neck Surgery,Chinese PLA Institute of Otolaryngology,Chinese PLA General Hospital,National Clinical Research Center for Otolaryngology Head and Neck Surgery,Key Laboratory of Hearing Impairment Science(Chinese PLA Medical School),Ministry of Education,Beijing Key Laboratory of Hearing Impairment Prevention and Treatment,Laboratory of Military Acoustic Trauma Protection,National Clinical Research Center for Geriatrics,Beijing 100853 China;Clinical Hearing Center,Affiliated Hospital of Xuzhou Medical University,Xuzhou 221006,China;Artificial Auditory Laboratory of Jiangsu Province,Xuzhou Medical University,Xuzhou 221004,China)

机构地区：[1]徐州医科大学听觉与平衡医学研究所,徐州221004 [2]首都医科大学附属北京友谊医院耳鼻咽喉头颈外科,北京100000 [3]中国人民解放军总医院耳鼻咽喉头颈外科,解放军总医院耳鼻咽喉研究所,耳鼻咽喉头颈外科国家临床研究中心,聋病教育部重点实验室,聋病防治北京市重点实验室,军事声损伤防护实验室,老年共病国家临床研究中心,北京100853 [4]徐州医科大学附属医院临床听力中心,徐州221006 [5]江苏省人工听觉重点实验室,徐州221004

出　　处：《中华耳科学杂志》2022年第3期445-451,共7页Chinese Journal of Otology

基　　金：国家自然科学基金(81800916);江苏省高校自然科学研究重大项目(19KJA560002);徐州市自然科学基金(KC20177);国家973计划国家重大科学研究计划干细胞项目(2012CB967900)。

摘　　要：目的研究氨基糖苷类抗生素在耳蜗内外毛细胞中的分布特征,以及这种分布方式对毛细胞功能可能产生的影响。方法采用荧光分子Texas Red标记庆大霉素,以100mg/kg浓度将荧光标记庆大霉素复合分子给予小鼠腹腔注射,持续给药14天,以正常的未给药小鼠作为对照组,分别于给药14天以及停止给药14天(28天)检测小鼠听力水平及庆大霉素在毛细胞中的分布情况。结果与对照组相比,庆大霉素持续暴露14天组ABR结果显示听力阈值有明显上移,共聚焦结果显示荧光标记庆大霉素在外毛细胞中分布于外毛细胞核上区接近纤毛的颈部区域,在内毛细胞核上区及核下区均有明显分布,在内毛细胞胞浆中呈瀑布状,特别是在内毛细胞底部带状突触区域有较为集中的荧光聚集。在停止注射庆大霉素14天后检测,听力并未得到改善且庆大霉素在内外毛细胞的分布情况并未发生显著变化,且毛细胞数量及排列未发生显著差别。结论庆大霉素暴露后可被耳蜗毛细胞显著摄取,其在内毛细胞中分布范围更加广泛,并且摄取的庆大霉素可在毛细胞内长时间存在,但毛细胞数量及排列未受明显影响。Objective To report distribution characteristics of aminoglycoside antibiotics in cochlear inner and outer hair cells and possible influence on hair cell function.Methods Gentamicin(100 mg/kg)was labeled with fluorescent molecule Texas Red and injected intraperitoneally daily in mice for 14 days.Normal control mice were injected with normal saline.After 14 and 28 days,distribution of fluorescent labeling in cochlear hair cells was observed by laser confocal microscope and auditory brainstem responses(ABRs)were used to estimate functional state of the cochlea.Results Compared with the control,ABRs showed significantly elevated thresholds after gentamicin exposure.Fluorescent labeled gentamicin was visible in the neck area near stereocilia in the outer hair cell nucleus,and widely distributed in the cytoplasm of the inner hair cells with a waterfall pattern.In particular,there was concentrated fluorescence aggregation in the ribbon synaptic region at the bottom of inner hair cells.At 14 days after treatment,there was no significant hearing threshold change and no significant change in gentamicin distribution in inner and outer hair cells,with no significant difference in the number and arrangement of hair cells compared to the control.Conclusion Upon continuous exposure,gentamicin can be significantly absorbed by cochlear hair cells and widely distributed in inner hair cells,which can last for a long time.However,the number and arrangement of hair cells are not significantly affected.

关 键 词：耳毒性 听力损失 庆大霉素 Texas Red 

分 类 号：R764[医药卫生—耳鼻咽喉科]