Molecular biological investigation for the treatment of inner ear disorders by controlling the free radicals t Medical

通过控制自由基治疗内耳疾病的分子生物学研究

基本信息

批准号：
10470357
负责人：
TAKUMIDA Masaya
金额：
$ 7.87万
依托单位：
Hiroshima University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10470357/
关键词：
inner ear NOS free radical nitric oxide fluorescence pathology molecular biology 蛍光 NO 内耳障害スーパーオキサイドメニエール病治療

项目摘要

Involvement of free radicals and nitric oxide (NO) in the certain inner ear disorders was investigated in order to found a way to study the mechanisms of inner ear disease. The localization of NO synthase (NOS) isoforms and free radicals were examined in inner ear of the normal and pathological guinea pig by immunohistochemistry. Production of NO in the isolated utricle or in the isolated vestibular hair cells after stimulation with L-arginine, glutamate, and gentamicin was investigated using DAF-2DA. Immunohistochemical stuandy demonstrated the expression for NOS II activity in the pathological inner ear. Production of NO in the inner ear was investigated using the fluorescence indicator, DAF-2DA for direct detection of NO. The fluorescence intensity of the sensory cells was augmented by stimulation with L-arginine, glutamate and gentamicin. The significant increase in NO production in the pathological animals was also noticed. These findings imply that NO from cNOS may mediate ototoxicity in the early phase whereas NO from NOS II may contribute to the late phase of tissue damage in the inner ear. For the aim to develop the new treatment of inner ear disorders, we performed a trial to assess the efficacy of the free-radical scavenger, rebamipide, in the treatment of Meniere' s disease and cisplatin-induced ototoxicity. Over 50% patients showed improvement of vertigo, healing disorders, tinnitus and disability in the Meniere's disease. Over 80% patients showed improvement of hearing in the cisplatin-induced ototoxicity. It has been suggested that treatment by controlling free radicals has a possibility to be an effective new therapy for inner ear disorders.These results were presented at meetings (8th and 9th meetings of Otological Society of Japan 57th and 58th meetings of Japan Society for equilibrium Research, 100th meeting of Oto-Rhino-Laryngological Society of Japan) and were presented by 12 papers .

研究了自由基和一氧化氮（NO）在某些内耳疾病中的参与，以找到一种研究内耳疾病机制的方法。通过免疫组织化学，在正常和病理豚鼠的内耳中检查了无合酶（NOS）同工型和自由基的定位。使用DAF-2DA研究了用L-精氨酸，谷氨酸和庆大霉素刺激后，在分离的肠内或分离的前庭毛细胞中的NO产生。免疫组织化学的Stuandy证明了病理内耳中NOS II活性的表达。使用荧光指标，DAF-2DA研究了内耳中的NO的产生，以直接检测NO。通过用L-精氨酸，谷氨酸和庆大霉素刺激增强感觉细胞的荧光强度。还注意到病理性动物的无生产的显着增加。这些发现表明，来自CNO的NO可以在早期阶段介导耳毒性，而NOS II的NO可能会导致内耳内耳损伤的晚期阶段。为了开发内耳疾病的新治疗方法，我们进行了一项试验，以评估自由基清除剂Rebamipide在治疗Meniere疾病和顺铂诱导的耳毒性方面的功效。超过50％的患者表现出眩晕，愈合障碍，耳鸣和脑膜病中的残疾改善。超过80％的患者在顺铂诱导的耳毒性中表现出听力的改善。有人认为，通过控制自由基的治疗可能是内耳障碍的有效新疗法。这些结果在日本大肠耳学学会的第57和第9届会议上提出了日本平衡研究协会第57和第58届会议，日本欧托 - 里希诺 - 里希诺（Oto-Rhino）学会100次会议日本的会议。