谷氨酸损伤体外培养大鼠螺旋神经元中凋亡诱导因子的表达与分布

发布时间：2018-06-09 12:05

本文选题：螺旋神经元 + 谷氨酸　；参考：《听力学及言语疾病杂志》2014年06期

【摘要】：目的观察不同浓度谷氨酸(Glu)损伤螺旋神经元中凋亡诱导因子(apoptosis inducing factor,AIF)的表达与分布变化,探讨利用谷氨酸损伤体外培养螺旋神经元是否与AIF凋亡途径相关。方法取40只出生后0~3天SD仔鼠螺旋神经元行体外培养,所得培养细胞平均分为正常组和10mM、20mM、40mM Glu组,正常组仅给予正常SGNs培养液培养,其余三组分别加入相应浓度谷氨酸和SGNs培养液,培养48h后,应用光学显微镜、免疫荧光染色及实时荧光定量PCR方法观察四组中SGN细胞形态、SGN中AIF的分布及AIF、钙蛋白酶(calpain)、半胱氨酸天冬氨酸3(caspase 3)的表达变化;用TUNEL法观察细胞凋亡。结果 20mM和40mM Glu组SGNs细胞形态改变和细胞凋亡明显,并伴有AIF核转位;不同浓度Glu组AIF和calpain基因表达明显高于正常组(P0.05),但caspase 3表达无明显升高(P0.05)。结论谷氨酸损伤体外培养的螺旋神经元过程中,AIF凋亡途径参与了细胞凋亡,calpain的高表达也促进了兴奋性神经递质对SGNs的损伤,但caspase3途径无明显作用。
[Abstract]:Objective to investigate the expression and distribution of apoptosis inducing factor-AIF-induced apoptosis in spiral neurons injured by glutamate at different concentrations, and to explore whether it is related to the apoptosis pathway of spiral neurons cultured with glutamate injury in vitro. Methods the spiral neurons of 40 Sprague-Dawley rats were cultured in vitro. The cultured cells were divided into normal group (n = 10) and control group (n = 10). The normal group was treated with SGNs culture medium only. The other three groups were incubated with glutamic acid and SGNs respectively for 48 hours. Immunofluorescence staining and real-time fluorescence quantitative PCR were used to observe the distribution of AIF and the expression of AIF, calpainin and caspic acid 3(caspase 3 in SGN cells in four groups, and apoptosis was observed by Tunel method. Results the morphological changes and apoptosis of SGNs cells in 20mm and 40mm Glu groups were obvious, and the expression of AIF and calpain genes were significantly higher in 20 mm and 40 mm Glu groups than in the normal control group, but the expression of caspase 3 was not significantly increased. Conclusion the high expression of calpain and the injury of excitatory neurotransmitters to SGNs may be involved in the apoptosis of spiral neurons injured by glutamate in vitro, but the caspase3 pathway has no obvious effect.
【作者单位】：第四军医大学西京医院耳鼻咽喉头颈外科;
【基金】：国家自然科学基金(81120108008,30930098,81271069) 国家“973”项目课题(2011CB504505)联合资助
【分类号】：R764

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