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目的观察鼓室内注射庆大霉素后,不同时间庆大霉素在前庭和耳蜗中的分布。方法将庆大霉素同德州红连接形成庆大霉素-德州红耦联物后,行豚鼠鼓室内注射,注射后12h,1、2、3、4、7、14、28d处死动物,Phalloidin染色后运用激光共聚焦扫描显微镜观察基底膜、椭圆囊、球囊、外半规管壶腹嵴庆大霉素分布情况,并进行荧光分布半定量分析。结果庆大霉素自注射后12h起在内耳所有细胞均见分布,在基底膜的外毛细胞、椭圆囊、球囊、外半规管壶腹嵴的感觉细胞聚集明显,主要聚集在毛细胞顶端纤毛下方的细胞质中,支持细胞分布较少。注射后第3天庆大霉素在内耳聚集达到最高峰,并在毛细胞内聚集较长时间。结论庆大霉素-德州红耦联物是一个研究庆大霉素在内耳分布的良好的荧光探针,可用来检测庆大霉素的药代动力学和聚集机制。
Objective To observe the distribution of gentamicin in the vestibule and cochlea after gentamicin injection at different time points. Methods Gentamicin and dendritic red were connected to form gentamicin-dezhou red conjugate. After guinea pigs were injected into the tympanic cavity, animals were sacrificed at 12h, 1,2,3,4,7,14,28d after injection. Phalloidin After staining, the distribution of gentamicin in basal lamina, oval sac, balloon and lateral semicircular canals was observed by confocal laser scanning microscopy. Semi-quantitative analysis of fluorescence distribution was performed. RESULTS: Gentamycin was found in all cells in the inner ear at 12h after injection. The sensory cells in the outer hair cells, the oval sac, the balloon and the outer semicircular canal in the basilar membrane gathered significantly, mainly in the apical ciliated hair cells Below the cytoplasm, supporting cells are less distributed. On the third day after injection, gentamicin reached the peak in the inner ear and aggregated for a long time in the hair cells. Conclusions Gentamicin-Texas Red Closure is a good fluorescent probe to study the distribution of gentamicin in the inner ear. It can be used to detect the gentamicin pharmacokinetics and aggregation mechanism.