红景天苷通过降低氧化应激介导氨基糖苷类药物耳毒性的抑制作用  

The Inhibitory Effect of Salidroside on the Ototoxicity of Aminoglycoside Drugs by Reducing Oxidative Stress

作　　者：于姝媛 李浩楠 朱紫嘉 王苹 杜波[1] YU Shu-Yuan;LI Hao-Nan;ZHU Zi-Jia;WANG Ping;DU Bo

机构地区：[1]吉林大学白求恩第一医院耳鼻喉头颈外科,长春130021

出　　处：《中国听力语言康复科学杂志》2018年第4期246-249,共4页Chinese Scientific Journal of Hearing and Speech Rehabilitation

基　　金：吉林省自然科学基金项目(20150101171JC)

摘　　要：目的研究红景天苷对氨基糖苷类药物耳毒性的抑制作用,并探讨NRF2/ARE途径的作用机制。方法出生4天的Wistar大鼠耳蜗基底膜,随机分为4组。正常培养的耳蜗基底膜为对照组;0.2 mmol/L的庆大霉素(gentamin,GM)作用24 h为模型组;400μmol/L的红景天苷(salidroside,SA)预处理培养耳蜗基底膜3 h后加GM作用24 h为干预组;400μmol/L的SA作用24 h为红景天苷组。在培养终止后分别检测耳蜗组织丙二醛(malondialdehyde,MDA)含量、超氧化物歧化酶(superoxide dismutase,SOD)和还原型谷胱甘肽(glutatione,GSH)活性。Phalloidin染色观察内外毛细胞数目的变化,实时定量PCR法检测NRF2、血红素加氧酶-1(home oxygenase,HO-1)和NAD(P)H醌氧化还原酶1(NAD(P)H dehydrogenase quinone1,NQO1)m RNA表达量。结果 (1)GM作用24 h出现毛细胞缺失,计数中底回外毛细胞缺失率为(38.6±8.92)%,内毛细胞缺失率为(41.5±6.42)%。SA预处理组内外毛细胞缺失率分别为(9.76±3.34)%和(11.38±7.14)%,与GM作用组相比毛细胞存活明显增加(P<0.01)。(2)分别在GM作用耳蜗后6、12和24 h时间点检测MDA、SOD和GSH,结果显示,MDA含量在GM作用后6 h升高,12 h达到峰值,24 h略降低,但仍高于对照组。SOD活性和GSH活性在GM作用后开始下降,呈时间依赖性。SA预处理可以明显增加SOD活性和GSH活性,降低MDA含量。与GM作用组相比有统计学差异(P<0.01)。(3)RT-PCR结果显示,NRF2及其下游调控因子HO-1和NQO1,在加入庆大霉素后比正常组升高,单纯SA组和SA干预组表达量更高。结论 SA通过激活NRF2/ARE途径促进下游相关联靶蛋白转录合成,从而发挥抗氧化作用,减轻GM诱导的耳蜗毛细胞损伤。Objective To study the inhibitory of salidroside on the ototoxicity of aminoglycoside drugs and to explore the mechanism of NRF2/ARE pathway.Methods The basement membrane of the newborn Wistar rat were randomly dividied into 4 groups.The normal cultured basement membrane worked as the control group,and 0.2 m M gentamin（GM） treated for 24 h were model group;400 μmol/L salidroside（SA） pretreatment for 3 h and then GM treatment for 24 h were intervention group;400 μmol/L alone treatment were SA group. The activity of MDA,SOD and GSH in cochlear tissue were detected after the termination of culture.The m RNA level of NRF2,HO-1 and NQO1 were detected by real time quantitative PCR. The haie cells loss was evaluated by phalloidin staining.Results（1） It appeared hair cells loss after GM treatment,the rate of（38.6±8.92） % in out hair cells and（41.5±6.42）% in inner hair cells. In the pretreatment of SA group, the rate of hair cells loss was（9.76±3.34）% and（ 11.38 ±7.14）%,and the survival hair cells increased significantly（P〈0.01） compared with GM treatment group.（2） MDA level was increased at 6 h after GM treatment,12 h reached peak,and 24 h decreased slightly. SOC and GSH activity began to decline after GM treated,and showed time dependent course. SA pretreated can significantly increase the activity of SOD and GSH,and decreased the level of MDA.（P〈0.01）（3） The results of q PCR showed that the expression of NRF2 and its downstream regulatory factors,HO-1 and NQO1 were higher in the SA group and SA pretreatment group than the GM group. Conclusion SA can promote NRF2/ARE pathway and be play a role in antioxidant effect and reducing the insult of cochlear hair cells induced by GM.

关 键 词：耳毒性 红景天苷 活性氧 毛细胞 

分 类 号：R285.5[医药卫生—中药学]