3-MA及OPTN对稳转SOD1-G93A突变蛋白NCS-34细胞系中自噬通路蛋白的作用  

The effects of 3-MA and OPTN on autophagy pathway proteins in NCS-34 cell line stably transformed with SOD1-G93A mutant protein

作　　者：温迪[1] 王娟[1] 李媛媛[1] 王燕燕[1] 刘亚坤[1] Wen Di;Wang Juan;Li Yuanyuan;Wang Yanyan;Liu Yakun(Department of Neurology,the Second Hospital of Hebei Medical University,Shijiazhuang 050000,China)

机构地区：[1]河北医科大学第二医院神经内科,石家庄050000

出　　处：《脑与神经疾病杂志》2025年第4期235-239,共5页Journal of Brain and Nervous Diseases

基　　金：河北省自然科学基金青年科学基金项目(H2021206048);河北省医学科学研究课题计划(20240793)。

摘　　要：目的探索以自噬通路抑制剂3-甲基腺嘌呤(3-MA)及慢病毒(LV)为载体介导的视神经病变诱导反应蛋白(LV-OPTN)对稳转超氧化物歧化酶1(SOD1-G93A)突变蛋白的NCS-34细胞系自噬通路蛋白OPTN、p62、微管相关蛋白ⅠA/ⅠB轻链3(LC3)、TANK结合激酶1(TBK1)的变化。方法采用稳转SOD1-G93A突变蛋白的NCS-34细胞系,并分为3组:无干预对照组、3-MA处理组(自噬通路抑制剂)、3-MA处理后给予LV-OPTN组,同时对3组细胞分别进行免疫荧光染色,特异性标记自噬通路蛋白p62、LC3、TBK1及OPTN,统计分析各蛋白的荧光含量。结果与对照组相比,3-MA组细胞中自噬相关蛋白的荧光强度p62增多,而LC3、TBK1及OPTN均减少,与对照组相比差异有统计学意义(^(均)P<0.05);对于3-MA预处理后再给予LV-OPTN组,我们观察到自噬通路相关蛋白的荧光表达有一定程度的恢复,即p62减少,而LC3、TBK1及OPTN均增加,与3-MA组相比差异均有统计学意义(^(均)P<0.05)。结论3-MA抑制稳转SOD1-G93A突变蛋白的NCS-34细胞系中自噬蛋白的活性,而LV-OPTN重新激活自噬,逆转了3-MA诱导的自噬通路的抑制作用。Objective To explore the changes of autophagy pathway proteins OPTN,p62,LC3,and TBK1in NCS-34 cell lines stabilized with SOD1-G93A mutant protein by inducing optic neuropathy response protein(LV-OPTN)mediated by autophagy pathway inhibitor 3-MA and lentivirus.Methods The NCS-34 cell line with stable SOD1-G93A mutant protein was used and divided into three groups:non intervention control group,3-MA treatment group(autophagy pathway inhibitor),and LV-OPTN group after 3-MA treatment.Immunofluorescence staining was performed on each group of cells to specifically label autophagy pathway proteins p62,LC3,TBK1,and OPTN,and the fluorescence content of each protein was statistically analyzed.Results Compared with the control group,the fluorescence intensity p62 of autophagy related proteins in the 3-MA group increased,while LC3,TBK1,and OPTN decreased,showing statistical differences compared with the control group(^(all)P<0.05);for the LVOPTN group after 3-MA pretreatment,we observed a certain degree of recovery in the fluorescence expression of autophagy pathway related proteins,with a decrease in p62 and an increase in LC3,TBK1,and OPTN,all of which were statistically different from the 3-MA group(^(all)P<0.05).Conclusion 3-MA inhibits the activity of autophagy proteins in the NCS-34 cell line with stable conversion of SOD1-G93A mutant protein,while LV-OPTN reactivates autophagy,reversing the inhibitory effect of 3-MA induced autophagy pathway.

关 键 词：肌萎缩侧索硬化 超氧化物歧化酶1 神经元细胞系 3-甲基腺嘌呤 视神经病变诱导反应蛋白 自噬通路 

分 类 号：R741[医药卫生—神经病学与精神病学]