应激心肌细胞中NGFI-B转位至线粒体的氨基酸序列研究  

Amino Acids Sequence of NGFI-B Translocation to Mitochondria in Cardiomyocytes of Stressed Rats

作　　者：冯红[1] 刘卫[2] 张德荣[1] 徐莉[1] 战锐[3] 王新兴[3] 钱令嘉[3] 

机构地区：[1]天津体育学院,天津300381 [2]军事医学科学院卫生学环境医学研究所,天津300050 [3]军事医学科学院基础所,北京100850

出　　处：《临床医学工程》2012年第5期702-705,共4页Clinical Medicine & Engineering

基　　金：国家自然科学基金资助项目(30370586)

摘　　要：目的探讨应激心肌细胞中NGFI-B发生线粒体转位的必需氨基酸序列,为揭示应激机体心肌损伤的病理学机理提供科学依据。方法建立心肌细胞应激损伤的离体细胞模型;构建NGFI-B缺失突变体与GFP的融合表达载体,分别命名为Δ1-pEGFP-N1(仅有N端152个AA的NGFI-B)和Δ2-pEGFP-N1(缺失N端152个AA的NGFI-B),并导入心肌细胞,采用细胞免疫荧光法观察其在应激心肌细胞中的亚细胞定位,采用流式细胞术观察离体心肌细胞的凋亡率。结果应激条件下,Δ1-pEGFP-N1转染心肌细胞后,发生向线粒体的转位,Δ2-pEGFP-N1转染心肌细胞后,仍滞留于核。Δ1-pEGFP-N1转染心肌细胞后,能显著降低应激引起的心肌细胞凋亡,Δ2-pEGFP-N1转染心肌细胞后,对应激引起的心肌细胞凋亡没有显著的保护作用。结论 NGFI-BN端的152个氨基酸对应激心肌细胞中NGFI-B发生线粒体转位进而导致细胞凋亡发挥重要作用。Objective To investigate the essential amino acids sequence of nerve growth factor-induced protein-B （NGFI-B） translocation to mitochondria in cardiomyocytes of stressed rats and to provide scientific evidences for exploring the mechanism underlying myocardium injury induced by stress. Methods The cell model of stress-induced cardiomyocyte injury was established. The GFP-tagged NGFI-B deletion mutants, named AI-pEGFP-N1 （constaining 152 AA from N-terminal） and A2-pEGFP-N1 （deleting 152 AA from N-terminal） respectively, were constructed and transfected into cardiomyocytes. The confocal microscopy method was used to investigate the subcellular location of NGFI-B deletion mutants in cardiomyocytes under stress. The flow cytometry was selected to detect the apoptotic rate in cardiomyocytes in vitro. Results Stress induced the translocation of AI-pEGFP-N1 from the nucleus to the mitochondria. Stress did not induce the translocation of A2-pEGFP-N1. Stress-induced cardiomyocyte apoptosis was decreased by AI-pEGFP-N1 transfection but not decreased by A2-pEGFP-N1 transfection. Conclusions 152 amino acids from N-terminal is the essential sequence ofNGFI-B translocating from the nucleus to the mitochondria and then resulting in cardiomyocyte apoptosis.

关 键 词：应激 心肌细胞 线粒体 NGFI-B 转位 氨基酸序列 

分 类 号：R-33[医药卫生]