摘要
目的利用斑马鱼模型研究三种抗真菌药物-氟康唑、联苯苄唑、制霉菌素-的耳毒性。方法1.分别探索三种药物的最大非致死浓度(MNLC)和10%的致死浓度(LC10);2.每种药物分别选取LC10、MNLC、1/3 MNLC和1/10 MNLC四个浓度进行研究。利用荧光立体显微镜和解剖显微镜观察并分析斑马鱼平衡囊、躯体形态及毛细胞数量的改变。5μg/mL的庆大霉素为阳性对照组,0.1%DMSO为阴性对照组,养鱼用水为空白对照组。结果1.氟康唑、联苯苄唑、制霉菌素三种药物的MNLC分别是653.8μg/mL、0.37μg/mL和10.5μg/mL。LC10分别是705.4μg/mL、0.52μg/mL和11.8μg/mL。2.三种药物均能导致斑马鱼平衡囊形态改变及身体屈曲。在LC10、MNLC、1/3 MNLC和1/10 MNLC四种浓度下,氟康唑组斑马鱼毛细胞损伤率分别为99%、95%、30%和28%;联苯苄唑组分别为94%、87%,、63%和49%;制霉菌素组分别为99%、89%、83%和25%。庆大霉素组斑马鱼毛细胞损伤率为55%。当与空白对照组比较,差异均具有统计学意义(P<0.001)。结论三种抗真菌药物均具有斑马鱼耳毒性,导致斑马鱼平衡囊形态发生改变并导致斑马鱼身体屈曲。三种药物均导致斑马鱼毛细胞数量的减少,并存在剂量依赖关系。
Objective The purpose of this research is to investigate ototoxicity of antifungal drugs(fluconazole,bifonazole,fungicidin)in zebrafish embryo and larvae.Methods Maximum non-lethal concentration(MNLC)and LC10 levels of the antifungal drugs were determined and four concentrations(LC10,MNLC,1/3 MNLC and 1/10 MNLC)were used to evaluate ototoxicity effects including abnormity in statocyst,flexed larvae trunk and hair cell loss.Gentamicin(5μg/mL)was used for positive control,fish water for untreated control,and DMSO(0.1%)for vehicle control.Images were acquired under a fluorescent stereomicroscope and a dissecting microscope and analyzed using the image J software.Results The MNLC was 653.8μg/mL,0.37μg/mL and 10.5μg/mL and the LC10 was 705.4μg/mL,0.52μg/mL and 11.8μg/mL for fluconazole,bifonazole and fungicidin,respectively.All three drugs caused changes in statocyst structures and trunk bending at resting state.Hair cell loss was seen at 99%,95%,30% and 28% in zebrafish treated with fluconazole,at 94%,87%,63% and 49% in those treated with bifonazole and at 99%,89%,83% and 25% in those treated with fungicidin at LC10,MNLC,1/3 MNLC and 1/10 MNLC,respectively.These were significantly different from those in untreated controls(P<0.001).Conclusion The three antifungal drugs can lead to ototoxicity in zebrafish affecting statocyst structures and causing trunk bending,as well partial or complete hair cell loss in statocyst and lateral line neuromasts,in a dose dependent manner.
作者
谌晶晶
徐慧芳
唐旭霞
CHEN Jingjing;XU Huifang;TANG Xuxia(Zhejiang Provincial Hospital of TCM,Hangzhou,310006)
出处
《中华耳科学杂志》
CSCD
北大核心
2021年第2期285-290,共6页
Chinese Journal of Otology
基金
浙江省卫生厅2015KYB266。
关键词
抗真菌药
斑马鱼
耳毒性
Ototoxicity
Zebrafish
Antifungal drugs
