AMPA受体在新生大鼠耳蜗螺旋神经节神经元兴奋毒性中的表达

Expression of AMPA Receptor in Excitotoxicity of Spiral Ganglion Neurons in Neonatal Rat Cochlea

叶文华;陈慧英;韦芳玉;林晓宇;刘金兰;易健启;叶莞丽;苏纪平;

1:广西医科大学第一附属医院耳鼻咽喉头颈外科

摘要

目的观察水杨酸钠(sodium salicylate, SS)作用新生SD大鼠耳蜗螺旋神经节神经元(spiral ganglion neuron, SGN)后,SGN中α-氨-3-羟基-5-甲基-4-异恶唑丙酸受体(α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors, AMPAR)在分子水平的表达改变。方法实验动物为10只新生3天的SD大鼠,分为免疫荧光染色组、正常对照组、水杨酸钠组、AMPAR激动剂组及AMPAR拮抗剂组,每组2只大鼠;采用免疫荧光染色技术检测各组大鼠SGN中AMPA受体GluA2的表达;采用FQ-PCR技术检测SGN和大脑皮层神经元中AMPAR各亚单位mRNA的表达;采用Western Blot技术检测SGN中GluA2的蛋白表达。结果免疫荧光染色显示,新生SD大鼠SGN胞体和轴突上均有AMPA受体GluA2的表达。FQ-PCR发现,①SGN及大脑皮层神经元中均有AMPAR亚单位GluA1、GluA2、GluA3和GluA4的表达(简称A1、A2、A3、A4),其中SGN中AMPAR各亚单位之间表达量呈A3>A2,A3>A4,A2和A4>A1。在大脑皮层神经元中,AMPAR各亚单位之间表达量呈A2>A3>A1>A4。②0.5 mM SS作用SGN后与正常对照组比较,只有A2在30 min时表达量明显降低(P<0.05)。③5 mM SS作用SGN后与正常对照组比较,A1～A4在30 min时表达量均降低(P<0.05);A2和A3表达量在1、3 h时降低(P<0.05);A2、A3和A4表达量在6 h时均降低(P<0.05)。Western Blot结果显示,5 mM SS作用SGN 30 min后与正常对照组比较,A2的膜蛋白表达量显著降低(P<0.05),总蛋白表达量变化无统计学差异(P>0.05)。结论①新生SD大鼠SGN及大脑皮层神经元中均有AMPAR表达,AMPAR各亚基在SGN及大脑皮层神经元中表达量高低变化呈不同规律。②水杨酸钠致SGN兴奋毒性中,与正常对照组比较,AMPAR的A2、A3和A4 mRNA表达量均显著降低,A2膜蛋白表达量亦明显下降;推测在水杨酸钠致SGN兴奋毒性中,水杨酸钠能促进A2膜蛋白表达量下调,促进AMPAR发生内化。

关键词

新生大鼠;水杨酸钠;耳蜗螺旋神经节神经元;兴奋毒性;α-氨-3-羟基-5-甲基-4-异恶唑丙酸受体

基金项目(Foundation):

国家自然科学基金项目(81560174,81360157);; 广西壮族自治区自然科学基金项目(2018GXNSFAA281173)资助

作者

叶文华;陈慧英;韦芳玉;林晓宇;刘金兰;易健启;叶莞丽;苏纪平;

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叶文华陈慧英韦芳玉林晓宇刘金兰易健启叶莞丽苏纪平