谷氨酸受体在噪声所致豚鼠螺旋神经节细胞损伤中的作用

Role of glutamate receptors in the spiral ganglion neuron damage induced by acoustic noise

本文旨在研究谷氨酸及其受体在噪声致豚鼠螺旋神经节细胞损伤中的作用。实验分为在体和离体两部分。(1)在体实验:豚鼠分为生理盐水(NS,10μL)组,NS(10μL)+噪声组和犬尿喹啉酸(kynurenic acid,KYNA,5 mmol/L,10μL)+噪声组,每组15只。用微量注射器经完整圆窗膜表面给予NS或KYNA;暴露于白噪声110 dB SPL,1 h。在圆窗给药前及噪声暴露后测试听觉脑干诱发电位(auditory brainstem response,ABR)阈值及Ⅲ波幅值,听神经复合动作电位(compound action potential,CAP)阈值及N1波幅值和潜伏期,测试后取基底膜进行透射电镜观察。(2)离体实验:观察高浓度谷氨酸对急性分离的豚鼠螺旋神经节细胞的影响。结果显示,NS+噪声组豚鼠ABR及CAP阈移显著高于KYNA+噪声组,且Ⅲ波和N1波幅值明显降低,潜伏期明显延长。NS+噪声组豚鼠毛细胞及传入神经末梢急性水肿和线粒体结构破坏;KYNA +噪声组豚鼠的毛细胞和传入神经末梢无明显变化。离体胞外施加谷氨酸可引起螺旋神经节细胞逐渐出现水肿、变性,最后死亡。本实验提示,噪声暴露可引起豚鼠听功能损伤,毛细胞/传入神经突触的结构破坏和螺旋神经节细胞变性、死亡;这种损伤可能与噪声暴露引起谷氨酸的过度释放有关;谷氨酸通过其受体介导致使螺旋神经节细胞损伤,谷氨酸受体的广谱拮抗剂KYNA可减轻噪声对螺旋神经节细胞的损伤。

The aim of the present study was to investigate the role of glutamate receptors in the damage of spiral ganglion neurons （SGNs） induced by acute acoustic noise. This investigation included in vivo and in vitro studies. In vivo, kynurenic acid （KYNA）, a broad-spectrum antagonist of glutamate receptors, was applied to the round window of guinea pigs, and its protective effect was observed. The animals were divided into three groups： control （saline, 0.9%, l0 I.tL）, saline （0.9%, l0 I.tL） ＋ noise and KYNA （5 retool/L, l0 I.tL） ＋ noise. Saline and KYNA were applied to the round window membrane with a microsyringe. The animals were exposed to 110 dB SPL of white noise for 1 h. Hearing thresholds for auditory brainstem responses （ABRs） and compound action potentials （CAPs） in all animals were measured before and after treatment. The amplitudes of III waveform of ABR and NI waveform of CAP and the latency of NI waveform at different stimulation levels （intensity-amplitude and intensity-latency functions） were also measured. The cochleas were then dissected for transmission electron microscopy （TEM） after final electrophysiological measurement. In vitro, the SGNs of the normal guinea pigs were isolated and glutamate （100 μmol/L or 1 000 μmol/L） was added into the medium. The morphology of the SGNs was examined by light microscopy. In vivo results showed that the hearing function and morphology of the inner ear including hair cells and SGNs in the control group were normal. Compared with that in the control group the thresholds for ABR and CAP （click and tone burst） in saline ＋ noise group were elevated significantly. The input-output functions showed that the amplitudes of Ⅲ waveform of ABR and N1 waveform of CAP decreased and the latency of N1 waveform increased obviously. There was significant difference in the amplitude and latency between saline ＋ noise group and KYNA ＋ noise group （P〈0.05）. TEM indicated obvious swelling and vacuoles at the terminate of dendrites of SGNs in NS ＋ noise group. On the contrary, the afferent dendrites in KYNA ＋ noise group showed normal appearance without swelling and vacuoles. In vitro experiment showed that the isolated SGNs of guinea pigs obviously swelled and even died after application of 100 μmol/L or 1 000 μmol/L glutamate. These results suggest that noise exposure causes hearing impairment, damage of hair cells and hair cell/afferent synapse and death of SGNs. The antagonist of glutamate receptors provides protective effects against hearing loss and SGN damage. It is inferred that excessive release of glutamate from the inner hair cells induced by noise may be responsible for these damages. Glutamate receptors are involved in the degeneration and death of SGNs.