PARP-1介导的AIF通路在链霉素致内耳毛细胞凋亡中的作用及机制研究

项目名称： PARP-1介导的AIF通路在链霉素致内耳毛细胞凋亡中的作用及机制研究

项目编号： No.81200744

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 医学二处

项目作者： 张道宫

作者单位： 山东大学

项目金额： 23万元

中文摘要： 应用氨基甙类抗生素引起的耳聋及平衡障碍是耳科学治疗的难题。研究表明氨基甙类抗生素损伤内耳的主要机理是毛细胞凋亡，但具体机制不明。我们前期研究发现氨基甙类抗生素导致的大鼠前庭毛细胞凋亡中有AIF参与，但关于AIF通路激活的具体机制不明。有文献报道AIF介导的凋亡依赖于PARP-1和Calpain的活化。我们拟在前期研究的基础上，建立大鼠链霉素内耳损伤体内外模型，通过流式细胞术和TUNEL法观察内耳毛细胞凋亡，应用Real-time、Western blot检测AIF、PARP-1和Calpain表达，运用共聚焦显微镜观察AIF核转位，阐明PARP-1介导的AIF通路在链霉素致内耳毛细凋亡中的作用及具体机制；在此基础上，抑制PARP-1或Calpain的表达，观察对内耳毛细胞凋亡及AIF核转位的影响。本研究将从新的视角阐明氨基甙类抗生素耳毒性机制，并为药物性耳聋及平衡障碍治疗提供新的靶点。

中文关键词： 聚腺苷酸二磷酸核糖转移酶-1；凋亡诱导因子；链霉素；毛细胞；凋亡

英文摘要： Deafness and balance disorders caused by the application of aminoglycoside antibiotics is one of the most difficult problems in otology. Studies have shown that aminoglycoside cause apoptosis of inner ear hair cells, but the exact mechanism has not been elucidated. Our previous study found that apoptosis inducing factor (AIF) involved in the aminoglycoside-induced apoptosis in rat vestibular hair cells, yet, the exact mechanisms of the activation of AIF pathway are not clear. Studies have shown that AIF mediated apoptosis is dependent on PARP-1 and Calpain activation. On the basis of our previous researches, the present project is designed to set the model of streptomycin-induced inner ear damage rats at overall and cellular level and to detect apoptosis of inner ear hair cells by flow cytometry and TUNEL. The expressions of PARP-1,Calpain and AIF will be detected by real-time PCR and western-blot. The nuclear translocation of AIF will be detected by confocal microscopy.We want to clarify the PARP-1-mediated AIF pathway in streptomycin-induced apoptosis of inner ear hair cells. On this basis, we will further observe the change of apoptosis of inner ear hair cells through the interference of PARP-1 or Calpain expression. Our new research will lends insight into the etiology and mechanisms responsible for aminogly

英文关键词： PARP-1；apoptosis inducing factor；streptomycin；hair cell；apoptosis