万伟华,黄文俪,温露
WAN Weihua,HUANG Wenli,WEN Lu

• 1:广东药科大学药学院

摘要(Abstract)：

目的建立一种快速筛选抗耳毒性化合物的斑马鱼毛细胞损伤模型。方法将受精后5 d(5 dpf)的斑马鱼幼鱼置于庆大霉素(10,25,50,100μmol·L~(-1))和DMSO溶液(0.2%,0.5%)中孵育不同的时间。DASPEI染色后,荧光显微镜下对神经丘(SO1、SO2、O1、OC1)中毛细胞进行观察和计数。结果 10μmol·L~(-1)庆大霉素溶液中孵育6 h达到近IC50效果,且0.2%DMSO对斑马鱼的毒性低。结论斑马鱼毛细胞损伤模型建立成功,可用于快速大规模筛选抗耳毒性化合物。
OBJECTIVE To establish a model of hair cell damage for rapid fishing of anti-ototoxic compounds. METHODS Five days post-fertilization( 5 dpf) zebrafish larvae were exposed respectively to gradient concentrations of gentamicin( 10,25,50,100μmol·L~(-1)) and DMSO( 0.2%,0.5%) for different time periods.Then hair cells within the four neuromasts( SO1,SO2,O1,OC1) were observed by fluorescence microscopy and the number of the hair cells was counted after stained with DASPEI. RESULTS Incubation in 10 μmol·L~(-1)gentamicin solution for 6 h could nearly achieve the effect of IC50,and 0.2% DMSO had an effect on zebrafish with low toxicity. CONCLUSION The zebrafish model of hair cell damage is successfully established and can be used for drug screening in a large scale quickly.

关键词(KeyWords)： 听力损伤;斑马鱼;庆大霉素;抗耳毒性化合物
hearing loss;zebrafish;gentamicin;anti-ototoxic compounds

Abstract:

Keywords:

基金项目(Foundation):

作者(Author): 万伟华,黄文俪,温露
WAN Weihua,HUANG Wenli,WEN Lu

参考文献(References)：

• [1]赵壮.斑马鱼药源性耳毒模型的建立及应用[D].北京:北京协和医学院,2011.
• [2]刘汉强.硫酸庆大霉素不同途径给药耳毒性临床研究[D].四川:四川大学,2003.
• [3]Govaerts P J,Claes J,Van De Heyning P H,et al.Aminoglycoside-induced ototoxicity[J].Toxicology letters,1990,52(3):227-251.
• [4]Warchol M E.Cellular mechanisms of aminoglycoside ototoxicity[J].Current opinion in otolaryngology&head and neck surgery,2010,18(5):454-458.
• [5]Liu C,Mu Y,Du J.Application of the zebrafish in the research of life sciences[J].Chinese Bulletin of Life Sciences,2007,19(4):382.
• [6]孙超,张森,高立栓,等.斑马鱼疾病模型的研究简述[J].中国新药杂志,2016,25(20):2340-2343.
• [7]Ton C,Parng C.The use of zebrafish for assessing ototoxic and otoprotective agents[J].Hearing research,2005,208:79-88.
• [8]Chang-Chien J,Yen Y C,Chien K H,et al.The connexin 30.3 of zebrafish homologue of human connexin 26 may play similar role in the inner ear[J].Hearing research,2014,313(7):55-66.
• [9]Chiu L L,Cunningham L L,Raible D W,et al.Using the zebrafish lateral line to screen for ototoxicity[J].Journal of the Association for Research in Otolaryngology,2008,9(2):178-190.
• [10]Harris J A,Cheng A G,Cunningham L L,et al.Neomycin-induced hair cell death and rapid regeneration in the lateral line of zebrafish(Danio rerio)[J].Journal of the Association for Research in Otolaryngology,2003,4(2):219-234.
• [11]Pinto-Teixeira F,Viader-Llargués O,Torres-Mejía E,et al.Inexhaustible hair-cell regeneration in young and aged zebrafish[J].Biology open,2015,4(7):903-909.
• [12]Owens K N,Santos F,Roberts B,et al.Identification of genetic and chemical modulators of zebrafish mechanosensory hair cell death[J].PLo S Genet,2008,4(2):e1000020.
• [13]Esterberg R,Coffin A B,Ou H,et al.Fish in a dish:drug discovery for hearing habilitation[J].Drug Discovery Today:Disease Models,2013,10(1):23-29.
• [14]陈宝增,李凤超,康现江.实验室斑马鱼的繁育与养殖技术[J].河北渔业,2008,4:51-53.
• [15]陈楠,高雯,张磊,等.水温、p H值和光照对斑马鱼受精卵孵化率的影响[J].水产养殖,2016,37(7):27-31.
• [16]Uribe P M,Mueller M A,Gleichman J S,et al.Dimethyl sulfoxide(DMSO)exacerbates cisplatin-induced sensory hair cell death in zebrafish(Danio rerio)[J].Plos One,2013,8(8):553-559.
• [17]Chang J,Choi J,Rah Y C,et al.Sodium Selenite Acts as an Otoprotectant against Neomycin-Induced Hair Cell Damage in a Zebrafish Model[J].Plos One,2016,11(3):151-557.
• [18]Owens K N,Coffin A B,Hong L S,et al.Response of mechanosensory hair cells of the zebrafish lateral line to aminoglycosides reveals distinct cell death pathways[J].Hearing research,2009,253(1):32-41.