谷氨酸受体5在人难治性颞叶癫痫脑组织中的表达研究  

The expression of glutamate receptor 5 in brain tissue of human intractable temporal lobe epilepsy

作　　者：曾义军[1,2] 余水[2] 雷町[1] 张恒[1] 李劲梅[3] 周章明[2] 

机构地区：[1]四川大学华西医院神经外科,成都610041 [2]都江堰市医疗中心神经外科,四川都江堰611830 [3]四川大学华西医院神经内科,成都610041

出　　处：《华西医学》2017年第5期659-664,共6页West China Medical Journal

基　　金：国家自然科学基金(0040205401243)

摘　　要：目的通过测定谷氨酸受体5(glutamic receptor 5,GLUR5)在人难治性颞叶癫痫和马桑内酯诱导的恒河猴癫痫模型脑组织中的蛋白表达水平,探讨GLUR5与难治性癫痫发病机制的关系。方法选取2007年1月—2015年12月四川大学华西医院收治的54例难治性颞叶癫痫患者作为临床病例组,同期收治的43例因外伤、肿瘤或大面积脑出血并发脑疝行去颞叶皮质减压手术的患者作为临床对照组。采用荧光定量聚合酶链反应(polymerase chain reaction,PCR)和蛋白质印迹法检测所收集标本的GLUR5的m RNA水平和蛋白表达水平。通过蛋白质印迹法在经马桑内酯诱导的恒河猴颞叶癫痫模型的海马及颞叶组织中检测GLUR5蛋白表达水平,并与动物对照组比较。结果荧光定量PCR结果显示临床病例组与临床对照组颞叶组织的GLUR5 m RNA表达比值(R值)为0.262,差异无统计学意义(P>0.05);海马组织的GLUR5 m RNA表达R值为4.896,差异有统计学意义(P<0.05);GLUR5扩增曲线显示临床病例组较临床对照组海马组织中GLUR5 m RNA水平向上调节;扩增曲线显示临床病例组与临床对照组颞叶组织中GLUR5 m RNA水平无明显变化。GLUR5 PCR扩增产物电泳图显示扩增片段为161 bp。蛋白质印迹实验显示临床病例组颞叶组织GLUR5/actin灰度值为2.172±0.063,临床对照组为2.142±0.060,差异无统计学意义(P>0.05);临床病例组海马组织为2.548±0.509;临床对照组为1.584±0.415,差异有统计学意义(P<0.05)。目的蛋白条带结果显示GLUR5蛋白表达临床病例组颞叶组织与临床对照组比较差异无统计学意义(P>0.05);而在海马组织中的表达明显高于临床对照组,差异有统计学意义(P<0.05)。恒河猴动物实验组海马组织GLUR5/actin灰度值为1.007±0.034,动物对照组为1.001±0.032,差异无统计学意义(P>0.05)。动物实验组颞叶组织GLUR5/actin灰度值为0.763±0.026,动物对照组为0.742±0.034,差异无统计学意义(P>0.05)。目Objective To discuss the correlation between glutamate receptor 5 （GLURS） and the pathogenesis of intractable temporal lobe epilepsy （ITLE）, through detecting the GLUR5 expression in human with ITLE and Coriaria lactone-induced rhesus monkey temporal lobe epilepsy model. Methods Fifty-four patients with ITLE treated in West China Hospital between January 2007 and December 2015 were regarded as clinical case group in this study. The other 43 patients who underwent temporal lobe removal decompression surgery in the same time period due to trauma, tumor or large area cerebral hemorrhage complicated with cerebral hernia were designated as the clinical control group. Quantitive polymerase chain reaction （PCR） and Western blot methods were used to detect mRNA and protein levels of GLURS. Western blot was also used to detect the GLUR5 protein level in the hippocampus and temporal lobe tissues of Coriaria lactone-induced rhesus monkey epilepsy model, and the result was compared with that of animal controls. Results Quantitive PCR results showed that the expression ratio （R value） of GLUR5 in the temporal lobe of the clinical case group to the clinical control group was 0.262, without significant difference （P〉0.05）, while the R value in the hippocampus was 4.896, with a significant difference （P〈0.05）. The amplification curve showed that the GLUR5 level in the hippocampus of the clinical case group was higher than that of the clinical control group, but the GLUR5 mRNA level in the temporal lobe tissue was not significantly changed. GLUR5 PCR amplified product electrophoresis showed that the amplified fragment was 161 bp. Western blot analysis showed that the GLURS/actin value of the temporal lobe tissue in the clinical case group was 2.172＋0.063, while the value in the clinical control group was 2.142±0.060, and the difference was not statistically significant （P〉0.05）. The GLUR5/actin value of the hippocampus in the clinial case group was 2.548±0.509, while it was 1.584±0.415 in th

关 键 词：红藻氨酸受体 谷氨酸受体5 难治性颞叶癫痫 恒河猴 

分 类 号：R742.1[医药卫生—神经病学与精神病学]