地塞米松对顺铂致斑马鱼毛细胞损伤的保护作用研究  

Protective Effects of Dexamethasone on Cisplatin-induced Hair Cell Injury in Zebrafsih

作　　者：涂钰莹 易英 涂玉梅 龙孝斌[1] Tu Yuying;Yi Ying;Tu Yumei;Long Xiaobin(Otolaryngology Head and Neck Surgery,Zhujiang Hospital,Southern Medical University/The Second School of Clinical Medicine,Southern Medical University,Guangzhou,510000,China)

机构地区：[1]南方医科大学珠江医院耳鼻咽喉头颈外科,广州510280

出　　处：《听力学及言语疾病杂志》2022年第1期64-68,共5页Journal of Audiology and Speech Pathology

摘　　要：目的探讨地塞米松对顺铂致斑马鱼侧线神经丘毛细胞损伤的保护作用。方法①将受精后5天(5 dpf)斑马鱼幼鱼分为对照组、顺铂组、顺铂+地塞米松组,每组10条。对照组置于系统水中;顺铂组斑马鱼幼鱼中加入50μmol/L顺铂孵育24小时;顺铂+地塞米松组预先给予25μmol/L地塞米松作用24小时,再给予50μmol/L顺铂孵育24小时;分别在移除顺铂后6、12、24、48 h时采用FM1-43FX染色标记斑马鱼侧线毛细胞,在荧光显微镜下检测三组毛细胞数。②利用顺铂、地塞米松建立斑马鱼氧化应激模型,在移除顺铂6 h后利用DCFH-DA、DPPP荧光分子探针测定三组斑马鱼幼鱼体内的活性氧(ROS)、脂质过氧化程度(MDA)。③采用实时荧光定量PCR(qPCR)检测三组抗氧化物酶基因sod1、cat1和gstr的表达水平。结果在移除顺铂后6、12、24 h,与对照组(圴为12.09±1.32个)相比,顺铂组(分别为2.38±0.65、4.45±0.61、6.37±1.08个)、顺铂+地塞米松组(分别为5.25±1.02、5.66±0.71、8.56±1.26个)侧线神经丘毛细胞数目明显降低(圴为P<0.05),地塞米松组与顺铂组相比差异均有统计学意义(均为P<0.05);而移除顺铂后48 h,三组的差异无统计学意义(P>0.05)。顺铂组斑马鱼体内ROS、MDA含量较对照组升高,差异均有统计学意义(均为P<0.05),同时,顺铂+地塞米松组斑马鱼体内ROS含量显著低于顺铂组(P<0.05)。顺铂组、顺铂+地塞米松组斑马鱼体内的cat1基因表达水平较对照组显著性下降(P<0.05),而三组间sod1、gstr基因表达差异无统计学意义(均为P>0.05)。结论地塞米松可以减弱顺铂导致的斑马鱼毛细胞耳毒性损伤,同时保护斑马鱼毛细胞免受氧化应激的影响。Objective To investigate the protective effect of dexamethasone on cisplatin induced injury of hair cells in the lateral line of zebrafish.Methods①The 5-day postfertilization(dpf)zebrafish larvae were divided into control group,cisplatin group and cisplatin+dexamethasone group.The larvac of zebrafish in control group were placed in the system water The larvae of zebrafish in cisplatin group were incubated with 50μmol/L cisplatin for 24 hours.The larvae in cisplatin+dexamethasone group were pretreated with 25μmol/L dexamethasone for 24 hours,and then incubated with 50μmol/L cisplatin for 24 hours.The hair cells of zebrafish lateral line were stained with FM1-43FX at 6,12,24 and 48 hours after the removal of cisplatin,and the+number of hair cells were measured under fluorescence microscope.②The oxidative stress model of zebrafish was established by cisplatin and dexamethasone.After removing cisplatin for 6 h,ROS and MDA in the larvae of three groups of zebrafish were measured by DCFH-DA and DPPP fluorescent molecular probes.③The expression levels of antioxidant enzyme genes sod1,cat1 and gstr were detected by real-time fluorescent quantitative PCR(qPCR).Results Compared with the control group,the number of hair cells in cisplatin group and cisplatin+dexamethasone group was significantly decreased at 6,12 and 24 h after cisplatin removal,while compared with cisplatin group,dexamethasone pretreatment group could alleviate the decrease of hair cell number(P<0.05).However,48 hours after cisplatin removal,there was no significant difference among the three groups.The contents of ROS and MDA in cisplatin group were significantly higher than control group(P<0.05),and the content of ROS in the cisplatin+dexamethasone group was significantly lower than that in the cisplatin group(P<0.05).The expression level of cat1 gene in zebrafish of cisplatin group and cisplatin+dexamethasone group was significantly lower than that of control group(P<0.05).Conclusion Dexamethasone can attenuate cisplatin induced ototoxicity and

关 键 词：斑马鱼毛细胞 顺铂 地塞米松 氧化应激 

分 类 号：R764.5[医药卫生—耳鼻咽喉科]