microRNA-183调节FOXO1对听觉毛细胞再生作用机制研究

The Mechanism of MicroRNA-183 Regulating FOXO1 on Auditory Hair Cell Regeneration

练东玲;涂钰莹;涂玉梅;龙孝斌

1:南方医科大学珠江医院耳鼻喉头颈外科/南方医科大学第二临床医学院

摘要
目的 探讨microRNA-183(miR-183)对斑马鱼内耳生长发育的影响,及其在顺铂损伤内耳神经丘毛细胞及耳囊、耳石后的再生和生长发育中的作用。方法 利用显微注射技术向斑马鱼受精卵内分别注入miRNA模拟物(agomir)或反义吗啉寡核苷酸(MOs),初步构建miR-183过表达或低表达模型,然后将受精后第72 h斑马鱼浸泡在50μmol/L顺铂溶液中24 h完成最终建模。利用miRNA微阵列分析、RT-qPCR等技术检测miR-183和FOXO1基因表达的改变;通过FM1-43FX荧光染色了解内耳神经丘毛细胞损伤及再生情况;使用正置显微镜观察耳囊、耳石生长发育变化。结果 (1)顺铂作用后斑马鱼内耳毛细胞受损且耳囊、耳石生长发育受阻;移除顺铂后内耳毛细胞可再生,耳囊、耳石生长发育也逐渐恢复。(2)miR-183在移除顺铂后被激活,是斑马鱼幼体miRNA中最有表达差异的成员之一(P<0.001),且表达量持续上调至峰值后逐渐恢复至正常水平。(3)与对照组相比,移除顺铂后过表达miR-183组可促进内耳毛细胞数量和耳囊耳石面积的恢复(P<0.01);低表达miR-183组则使其恢复受到抑制(P<0.05)。(4)移除顺铂作用后FOXO1被激活,表达量上调(P<0.01);当过表达miR-183时FOXO1受到抑制,表达下调(P<0.01);低表达miR-183时FOXO1则表达量上升(P<0.01)。结论 miR-183与移除顺铂作用后内耳毛细胞的再生、耳囊耳石面积生长发育的恢复呈正向调控关系,其机制可能与miR-183对FOXO1的负向调控有关。
关键词
斑马鱼;顺铂;毛细胞;MicroRNA-183;FOXO1
基金项目(Foundation):
作者
练东玲;涂钰莹;涂玉梅;龙孝斌
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