[1]姜宏佺,丰宏林,孙晓嘉.胍那苄对运动神经元的内质网应激细胞模型具有保护作用[J].卒中与神经疾病杂志,2017,24(03):173-176+196.[doi:10.3969/j.issn.1007-0478.2017.03.001]
　Jiang Hongquan,Feng Honglin,Sun Xiaojia..Protective effect of guanabenz in endoplasmic reticulum stress model of mouse motor neuron-like hybrid cell line[J].Stroke and Nervous Diseases,2017,24(03):173-176+196.[doi:10.3969/j.issn.1007-0478.2017.03.001]
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胍那苄对运动神经元的内质网应激细胞模型具有保护作用()
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《卒中与神经疾病》杂志[ISSN:1007-0478/CN:42-1402/R]

卷:
第24卷
期数:
2017年03期
页码:
173-176+196
栏目:
论 著
出版日期:
2017-06-26

文章信息/Info

Title:
Protective effect of guanabenz in endoplasmic reticulum stress model of mouse motor neuron-like hybrid cell line
文章编号:
1007-0478(2017)03-0173-05
作者:
姜宏佺; 丰宏林; 孙晓嘉
150001 哈尔滨医科大学附属第一医院神经内科疑难病科[姜宏佺 丰宏林(通信作者)孙晓嘉]
Author(s):
Jiang Hongquan; Feng Honglin; Sun Xiaojia.
Department of Neurology,the First Affiliated Hospital of Harbin Medical University,Harbin 150001
关键词:
神经系统变性病 肌萎缩侧索硬化 胍那苄 内质网应激 SOD1
Keywords:
Neurodegenerative disease Amyotrophic lateral sclerosis Endoplasmic reticulum stress Guanabenz SOD1
分类号:
R741
DOI:
10.3969/j.issn.1007-0478.2017.03.001
摘要:
目的 探讨P-eIF2α去磷酸化抑制剂胍那苄(guanabenz,GA)对胚胎小鼠运动神经元样杂交细胞系(mouse motor neuron-like hybrid cell line,NSC34)的内质网应激模型是否具有保护作用及是否能够减少外源性SOD1 G93A蛋白的合成。方法(1)应用依霉素(Tunicamycin,TM)作用到鼠运动神经元样杂交细胞系(mouse motor neuron-like hybrid cell line,NSC34),获得运动神经元ERS模型; 再给予不同浓度的GA通过显微镜及MTT方法观察GA对运动神经元ERS模型细胞数量及细胞活力的影响; 用Western Blot方法检测TM、GA对NSC34细胞系ERS蛋白eIF2α和P-eIF2α的影响;(2)应用慢病毒转染的方法制作SOD1 G93A NSC34稳定转染细胞系,通过Western blot方法检测GA对外源性SOD1 G93A蛋白以及内源性mouse SOD1蛋白表达量的影响。结果(1)在NSC34的ERS模型中多个浓度的GA与对照组比较可以显著增加细胞数量,提高细胞活力(P<0.05); GA可以增加ERS相关蛋白P-eIF2α的表达量;(2)GA在SOD1 G93A NSC34细胞系中能够减少外源性hSOD1蛋白的表达量,而对内源性的mouse SOD1蛋白表达量没有明显影响。结论(1)GA可能通过增加P-eIF2α的表达量而对NSC34细胞系的ERS模型具有保护作用;(2)GA在SOD1 G93A NSC34细胞系中可以减少外源性hSOD1蛋白的表达量。
Abstract:
ObjectiveTo explore the protective effect of guanabenz(GA),a novel inhibitor of eukaryotic initiation factor 2α(eIF2α)dephosphorylation,in endoplasmic reticulum stress(ERS)model of mouse motor neuron-like hybrid cell line(NSC34),and determine whether it could decrease the expression of exogenous hSOD1 G93A protein.Methods(1)Tunicamycin(TM)was used to stress NSC34 to produce ERS model.We tested the cell's number and viability of NSC34 treated with different concentrations of GA by microscope and MTT test.The expression of phosphorylated-eIF2α(P-eIF2α)and eIF2α was detected by western blot in Vehicle,TM and TM+GA groups.( 2)hSOD1 G93A was transfected into NSC34 with lentiviral.The expression of hSOD1 G93A and mouse SOD1 was detected by Western blot in hSOD1 G93A NSC34 treated with or without GA.Results GA increased the cell's number and viability of NSC34 treated with different concentrations of GA compared with control group(P<0.05).Western blot results revealed that GA dramatically increased the level of P-eIF2α protein,without affecting total eIF2α protein level.The exogenous hSOD1 G93A protein was decreased by GA,but the expression of mouse endogenous SOD1 was not affected.Conclusion GA can protect the ERS model of NSC34 through increasing the expression of P-eIF2α protein.GA can decrease insoluble mutant SOD1 aggregates.

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备注/Memo

备注/Memo:
基金项目:黑龙江省教育厅科学技术研究项目资助(编号为12541472)
更新日期/Last Update: 2017-06-20