氨基糖苷类药物耳毒性的斑马鱼模型的研究  

Zebrafish model for the study on drug ototoxicity of aminoglycoside antibiotics

作　　者：赵壮[1] 佟军威[1] 张靖溥[1] 游学甫[1] 蒋建东[1] 胡昌勤[2] 

机构地区：[1]中国医学科学院北京协和医学院医药生物技术研究所,北京100050 [2]中国食品药品检定研究院,北京100050

出　　处：《药学学报》2011年第8期928-935,共8页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金面上项目(30772681);国家"重大新药创制"科技重大专项(2009ZX09301-003-6-22008ZX09305-001)

摘　　要：氨基糖苷类抗生素因其抗菌谱广、抗菌能力强,半个多世纪以来一直是临床上常用的抗菌素之一。但氨基糖苷类抗生素具有很强的耳毒和肾毒作用,在药物致聋因素中排在首位。本研究以庆大霉素(gentamycin)、新霉素(neomycin)、链霉素(streptomycin)等3种氨基糖苷类抗生素为代表性药物,研究其对斑马鱼胚胎发育的毒性作用和对幼体毛细胞的损伤作用,并探索了该损伤与听觉相关基因之间的联系。结果显示:①3种药物的致死作用都具有明显的浓度依赖性,其致死作用的强弱顺序为链霉素>新霉素>庆大霉素;②3种药物处理的5 dpf(day past fertilization)幼体出现身体失衡及体位异常,以及耳囊结构的异常变化;③毛细胞染色实验可观察到,3种药物作用的毛细胞和神经丘均出现明显的损伤和数量减少;④与听觉器官发育相关的基因eya1、val、otx2、dlx6a均随3种抗生素药物浓度的升高,出现差异性的表达水平下调。本研究首次探索了这3种耳毒性氨基糖苷类抗生素处理与斑马鱼听囊结构和听觉基因表达的相关性;并证明利用斑马鱼建立简便、准确、直观、快速地检测药物耳毒性的模型和检测方法的可行性。Aminoglycoside antibiotics,due to their strong antibacterial effects and broad antimicrobial spectra,have been very commonly used in clinical practice in the past half century.However,aminoglycoside antibiotics manifest severe ototoxicity and nephrotoxicity,and are one of top factors in hearing loss.In this study,three members of the aminoglycoside antibiotics family,gentamycin,neomycin and streptomycin,were chosen as the representatives to be investigated for their toxicity to the embryonic development and the larva hair cells in zebrafish,and also to their target genes associated with hearing-related genes.The results showed that： ① the lethal effect of all three drugs demonstrated a significant dependence on concentration,and the severity order of the lethal effect was streptomycin neomycin gentamycin;② all the three drugs caused the larva trunk bending in resting state at 5 dpf（day past fertilization）,probably due to their ototoxicity in the physical imbalance and postural abnormalities;③ impairment and reducing of the hair cells were observed in all three cases of drug treatment;④ four genes,eya1,val,otx2 and dlx6a,which play an important role in the development of hearing organs,showed differential and significant decrease of gene expression in a drug concentration-dependent manner.This study for the first time reports the relevance between the expression of hearing genes and the three ototoxic antibiotics and also proved the feasibility of establishing a simple,accurate,intuitive and fast model with zebrafish for the detection of drug ototoxicity.

关 键 词：氨基糖苷类抗生素 耳毒性 斑马鱼模型 毛细胞 听力基因 

分 类 号：R963[医药卫生—微生物与生化药学]