肌萎缩性脊髓侧索硬化症细胞模型中mRAGE和esRAGE选择性剪接变化研究  

Study on alternative splicing changes of mRAGE and esRAGE in ALS cell model

作　　者：倪晶 庄菊花 王国玉 NI Jing;ZHUANG Ju-hua;WANG Guo-yu(Department of Nuclear Medicine,The Seventh People＇s Hospital,Shanghai University of Traditional Chinese Medicine,Shanghai 200137,China.)

机构地区：[1]上海中医药大学附属第七人民医院核医学科,上海200137

出　　处：《临床和实验医学杂志》2018年第18期1918-1921,共4页Journal of Clinical and Experimental Medicine

基　　金：上海市卫生和计划生育委员会青年项目资助(编号:20144Y0115);上海市浦东新区卫计委优秀青年医师培养计划项目资助(编号:PWRq2015-12)

摘　　要：目的探讨肌萎缩性脊髓侧索硬化症(ALS)细胞模型中晚期糖基化终末产物受体(RAGE)两种亚型mRAGE和esRAGE选择性剪接变化。方法选取鼠运动元样细胞系NSC-34细胞为研究对象,以SH-SY5Y细胞c DNA为模板,构建铜/锌超氧化物歧化酶(SOD1)突变载体。将NSC-34细胞分为:空质粒组、正常SOD1蛋白表达质粒组、G93A质粒组,分别将空质粒、正常SOD1蛋白表达质粒、SOD1突变蛋白表达质粒(G93A质粒组)转染入细胞,另以正常NSC-34细胞组作为对照,不转染。转染后48 h,观察各组细胞形态学变化,采用实时聚合酶链式反应(RT-PCR)检测SOD1、mRAGE、esRAGE表达情况。结果 G93A质粒组与正常NSC-34细胞组细胞形态学比较,细胞胞体变圆,轴突变短且突起减少,空质粒组、正常SOD1蛋白表达质粒组与正常NSC-34细胞组比较无明显变化;qRT-PCR结果显示,正常SOD1蛋白表达质粒组、G93A质粒组细胞中SOD1为阳性表达,空质粒组及正常NSC-34细胞组SOD1为阴性表达,提示成功构建ALS细胞模型;与正常SOD1蛋白表达质粒组比较,G93A质粒组细胞中mRAGE表达明显增加(P<0.05),esRAGE表达明显降低(P<0.05)。结论在ALS病理过程中,胶质细胞中RAGE的选择性剪接调控异常改变,导致mRAGE亚型增加,esRAGE亚型减少。Objective To explore alternative splicing changes in mRAGE and esRAGE two subtypes of advanced glycation end products receptor（RAGE） in amyotrophic lateral sclerosis（ALS） cell model. Methods NSC-34 cell of rat motor like cell line was selected as the study objects,and SH-SY5 Y cell c DNA was used as template,and copper/zinc superoxide dismutase（SOD1） mutant vector was structured. NSC-34 cells were divided into empty plasmid group,normal SOD1 protein expression plasmid group and SOD1 mutant protein expression plasmid（G93 A plasmid group）. The empty plasmid,the normal SOD1 protein expression plasmid,and the G93 A plasmid were transfected into the cells,respectively. What＇s more,the normal NSC-34 cell group without transfection was used as a control. 48 hours after transfection,the morphological changes of each group were observed,and the expression of SOD1,mRAGE and esRAGE was detected by real-time polymerase chain reaction（RT-PCR）. Results Cell morphology comparison between the G93 A plasmid group and normal NSC-34 cell group,cell bodies became rounded,axons shortened and the processes decreased. There were no significant changes among the empty plasmid group,the normal SOD1 protein expression plasmid group and the normal NSC-34 cell group. QRT-PCR result showed that SOD1 was positive-expression in the normal SOD1 protein expression plasmid group and the G93 A plasmid group,and SOD1 was negative-expression in the empty plasmid group and the normal NSC-34 cell group,indicating that ALS cell model was successfully constructed. Compared with the normal SOD1 protein expression plasmid group,the expression of mRAGE was significantly increased in the G93 A plasmid group（P〈0. 05）,and the expression of esRAGE was significantly decreased（P〈0. 05）. Conclusion Aberrant splicing regulation of RAGE in glial cells changes abnormally in the course of ALS pathology,it cause mRAGE subtype increased and esRAGE subtype decreased.

关 键 词：肌萎缩性脊髓侧索硬化症 晚期糖基化终末产物受体 mRAGE ESRAGE 

分 类 号：R744.8[医药卫生—神经病学与精神病学]