长期注射水杨酸钠对大鼠下丘GAD67和GABAAα1、c-fos表达的影响

doi:10.6040/j.issn.1673-3770.0.2014.330

摘要/Abstract

摘要： 目的通过观察长期注射水杨酸钠对大鼠听性脑干反应(ABR)及下丘GAD67和GABAAα1、c-fos表达的影响,探讨大鼠下丘GAD67、GABAAα1、c-fos表达的改变在水杨酸钠耳毒性中的可能机制。方法将24只成年健康Wistar大鼠随机分为2组:水杨酸钠组(肌肉注射10%水杨酸钠175 mg/kg,2次/d,连续28 d)、对照组(每天相同时间注射等量生理盐水,连续28 d)。两组大鼠在处死前进行ABR检测并观察ABR反应阈及波Ⅲ潜伏期的变化,然后将大鼠断头处死并迅速剥离下丘,采用实时荧光定量PCR(real-time PCR)、蛋白质印迹(Western blot)方法检测下丘GAD67、GABAAα1、c-fos mRNA及蛋白表达的变化。结果 ①药物注射28 d后,水杨酸钠组较注射前以及对照组ABR反应阈明显升高,波Ⅲ潜伏期明显延长,差异有统计学意义(P<0.01);②水杨酸钠组GAD67、GABAAα1、c-fos mRNA及其蛋白表达水平较对照组明显升高,差异有统计学意义(P<0.01)。结论 GAD67、GABAAα1、c-fos均不同程度参与了水杨酸钠耳毒性的发生发展过程, c-fos表达的上调则可能与水杨酸钠作用于机体后引起听觉中枢神经活动增强有关。

关键词: 水杨酸钠, 下丘, GAD67, GABAAα1, c-fos

Abstract: Objective To observe the influences of sodium salicylate on the auditory brainstem response(ABR)and expression of GAD67、GABAAα1、 c-fos in rat inferior colliculus. To explore the underlying mechanisms of these changes on sodium salicylate induced ototoxicity. Methods Twenty-four Wistar rats were randomly divided into two groups:sodium salicylate group(intramuscular injection of 10% sodium salicylate, 175 mg/kg, twice daily for 28 days)、control group(intramuscular injection with saline on same does at the same time).The rats were detected ABR and observe the changes of ABR threshold before they were killed,then the rats were decapitated and inferior colliculus tissue were stripped. Real-time reverse transcriptase polymerase chain reaction(real-time RT-PCR)and Western blot were used to study the different expression of GAD67、GABAAα1、c-fos mRNA and protein level in the two groups. Results ①ABR threshold and latency of wave Ⅲ of sodium salicylate group was increased and prolonged remarkably than that of before administration and the control after injection for 28 days(P<0.01);②The inferior colliculus GAD67、GABAAα1、c-fos mRNA and protein expression level of sodium salicylate group was significantly higher than the control group(P<0.01). Conclusion In the long-term effects of sodium salicylate, the upregulation of GAD67 and GABAAα1 expression may occur as a compensatory and regulatory mechanisms for sodium salicylate ototoxicity, the elevated expression of c-fos may be associated with the increased activity of central auditory system after sodium salicylate effects on the body.

Key words: Sodium salicylate, Inferior colliculus, GABAAα1, c-fos, GAD67

中图分类号:

R765.25

杨琨, 吴莎, 华清泉. 长期注射水杨酸钠对大鼠下丘GAD67和GABAAα1、c-fos表达的影响[J]. 山东大学耳鼻喉眼学报, 2015, 29(1): 24-27.

YANG Kun, WU Sha, HUA Qing-quan. Effect of sodium salicylate on the expression of c-fos, GAD67 and GABAAα1 in rat inferior colliculus[J]. JOURNAL OF SHANDONG UNIVERSITY (OTOLARYNGOLOGY AND OPHTHALMOLOGY), 2015, 29(1): 24-27.

参考文献

[1] Kojda G. Direct casoprotection by aspirin: a significant bonus to antiplatelet activity?[J].Cardiovasc Res, 2004, 64(2):192-194.
[2] Ramesh G, Reeves W B. Salicylate reduces cisplatin nephrotoxicity by inhibition of tumor necrosis factor-aloha[J]. Kidney Int, 2004, 65(2):490-499.
[3] Kinitsuki T, Kakazu Y, Matsumoto N, et al. Salicylate-induced morphlogical changes of isolated inner hair cells and outer cells from guinea-pig cochlea[J].Auris Nasus Larynx, 2009, 36(2):152-156.
[4] Guitton M J, Caston J, Reul J, et al.Salicylate induces tinnitus through activation of cochlear NMDA receptors[J]. J Neurosci, 2003, 23(9):3944-3952.
[5] Ruel J, Chabbert C, Nouvian R, et al. Salicylate enables cochlear arachidonic-acid-sensitive NMDA receptor responses[J]. J Neurosci, 2008, 28(29):7313-7323.
[6] Huang Z W, Luo Y, Wu Z, et al. Paradoxical enhancement of active cochlear mechanics in Long-term administration of salicylate[J]. J Neurophysiol, 2005, 93(4):2053-2061.
[7] Peleg U, Perez R, Freeman S, et al. Salicylate ototoxicity and its implications for cochlear microphonic potential generation[J]. J Basic Clin Physiol Pharmacol, 2007, 18(3):173-188.
[8] Eggemontt J J. Pathophysiology of tinnitus[J]. Prog Brain Res, 2007, 166:19-35. Review.
[9] 陈林,王海涛,陆云刚,等.水杨酸钠诱导耳鸣的神经机制[J].中国科学技术大学学报,2008,38(8):986-995.CHEN Lin, WANG Haitao, LU Yungang, et al. Neural mechanisms underlying tinnitus induced by sodium salicylate[J]. Chin J Univ Sci Tech, 2008, 38(8):986-995.
[10] Bauer C A, Brozoski T J, Holder T M, et al. Effects of chronic salicylate on GABAergic activity in rat inferior colliculus[J]. Hear Res, 2000, 147(1-2):175-182.
[11] Sun W, Lu J, Stolzberg D, et al. Salicylate increases the gain of the central auditory system[J]. Neuroscience, 2009, 159(1):325-334.
[12] Mahlke C, Wallhusser-Franke E. Evidence for tinnitus-related plasticity in the auditory and limbic system, demonstrated by arg3.1 and c-fos immunocytochemistry[J]. Hear Res, 2004, 195(1-2):17-34.
[13] 王洪田,孙建和,于宁,等.水杨酸钠引起耳鸣大鼠听皮层突触形态改变的透射电镜观察[J].中华耳科学杂志,2009,7(3):208-211. WANG Hongtian, SUN Jianhe, YU Ning, et al. Prolonged effects of salicylic acid on ultrastructure of auditory cortex synapses in a rat model of tinnitus[J]. Chin J Otology, 2009, 7(3):208-211.
[14] 陈抗松,廖华,杨希林,等.水杨酸致耳鸣大鼠的行为学及听性脑干反应改变[J].听力学及言语疾病杂志,2013,21(2):163-166. CHEN Kangsong, LIAO Hua, YANG Xilin, et al. Changes of the Behavior and ABR on the Tinnitus Rat Model Induced by Salicylate[J]. J Audiol Speech Pathol, 2013, 21(2):163-166.
[15] Sun W, Lu J, Stolzberg D, et al.Salicylate increases the gain of the central auditory system[J]. Neuroscience, 2009, 159(1):325-334.
[16] Wang H T, Luo B, Zhou K Q, et al. Sodium salicylate reduces inhibitory postsynaptic currents in neurons of rat auditory cortex[J]. Hear Res, 2006, 215(1-2):77-83.
[17] Burianova J, Ouda L, Profant O, et al. Age-related changes in GAD levels in the central auditory system of the rat[J]. Exp Gerontol, 2009, 44(3):161-169.
[18] Pouyatos B, Morel G, Lambert-Xolin A M, et al. Consequences of noise-or styrene-induced cochlear damages on glutamate decarboxylase levels in the rat inferior colliculus[J]. Hear Res, 2004, 189(1-2):83-91.
[19] Hwang I K, Li H, Yoo K Y, et al. Comparison of glutamic acid decarboxylase 67 immunoreactive neurons in the hippocampal CA1 region at various age stages in dogs[J]. Neurosci Lett, 2008, 431(3):251-255.
[20] Esclapez M, Houser C R.Up-regulation of GAD65 and GAD67 in remaining hippocampal GABA neurons in a model of temporal lobe epilepsy[J]. J Comp Neurol, 1999, 412(3):488-505.
[21] Zou Q Z, Shang X L. Effect of salicylate on the large GABAergic neurons in the inferior colliculus of rats[J]. Acta Neurol, 2012, 112(4):367-374.

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