庆大霉素致大鼠耳蜗氧化损伤的差异基因表达谱分析

Analysis of Differential Gene Expression Profile of Gentamicin-Induced Oxidative Damage in Rat Cochlea

杨阳;米文娟;田超永;林颖;查定军;

1:中国人民解放军空军军医大学西京医院耳鼻咽喉头颈外科全军内耳研究重点实验室

摘要
目的 研究庆大霉素损伤后耳蜗的顶、中和底回中与氧化应激相关的基因表达谱,以期筛选防治药物性聋的候选基因。方法 选取P2 SD大鼠15只,分离基底膜,将基底膜分为对照组与实验组,每组15条基底膜;实验组使用0.6 mM庆大霉素培养24小时,对照组使用等量PBS培养24小时;每组5条基底膜分别进行免疫荧光染色和毛细胞计数,对照组与实验组各10条基底膜,分为顶、中和底回各三组,提取RNA,利用RT2 Profiler2 Profiler(TM) PCR阵列筛选六组与氧化损伤相关的84个基因的表达差异,并对差异基因进行GO分析和相互作用网络分析;利用RT-PCR对候选基因进行验证。结果 实验组庆大霉素损伤后,毛细胞数量减少,且中回和底回的毛细胞数量显著少于顶回;与对照组相比,实验组中84个氧化损伤相关的基因中,顶回共有14个基因表达有生物学意义,其中12个基因表达上调,2个基因表达下调;中回共有19个基因表达有生物学意义,其中5个基因表达上调,14个基因表达下调;底回共有10个基因表达有生物学意义,其中5个基因表达上调,5个基因表达下调;其中Hmox1、Ptgs2、Apoe、Gpx6四个基因在庆大霉素损伤后差异倍数从顶回到底回逐渐减小。蛋白互作分析显示Hmox1是具有最高连接度的关键基因,与9个基因相互作用。RT-PCR验证显示Hmox1基因表达量在庆大霉素损伤后沿耳蜗纵轴差异倍数逐渐减小。结论 庆大霉素损伤过程中,Hmox1可能通过上调其在基底膜顶回的表达起到保护作用,Hmox1可作为庆大霉素损伤中起保护作用的关键候选基因进行深入研究。
关键词
庆大霉素;耳毒性;氧化损伤;基因表达差异
基金项目(Foundation):
国家重点基础研究发展规划项目(2014CB541700);; 国家自然科学基金项目(81870732、81670924);; 陕西省科技重点项目(2018PT-01);陕西省科技重点项目(2017ZDXM-SF-061);; 西京助推-先进学科建设项目(XJZT14X07、XJZT18ML18、XJZT19MDT02)联合资助
作者
杨阳;米文娟;田超永;林颖;查定军;
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