铁硫簇结合蛋白1的磁感应能力  

Ability of iron sulfur cluster binding protein 1 to respond to magnetic fields

作　　者：刘梦楠 田萧羽 方文灿 吴宁 李锦 李红 LIU Mengnan;TIAN Xiaoyu;FANG Wencan;WU Ning;LI Jin;LI Hong(State Key Laboratory of Toxicology and Medical Countermeasures,Academy of Military Medical Sciences,Beijing 100850,China;Medical School of Chinese PLA,Beijing 100853,China)

机构地区：[1]军事医学研究院国家安全特需药品全国重点实验室,北京100850 [2]中国人民解放军医学院,北京100853

出　　处：《中国药理学与毒理学杂志》2024年第6期420-425,共6页Chinese Journal of Pharmacology and Toxicology

基　　金：国家自然科学基金(82373853);国家自然科学基金(82202523)。

摘　　要：目的探究铁硫簇结合蛋白1(ISCA1)磁场刺激后对细胞钙内流的影响。方法(1)制备携带ISCA1基因的质粒、携带Magneto 2.0序列的质粒和空载质粒,且3种质粒均携带mCherry基因,包装成慢病毒感染HEK293A细胞,荧光显微镜观察慢病毒感染效率。(2)免疫共沉淀技术检测ISCA1与隐花色素1(CRY1)和CRY2蛋白之间的结合。(3)在磁场(40 mT,0.1 Hz,90%占空比)条件下,活细胞钙成像技术检测高表达ISCA1或Magneto 2.0细胞的钙内流。结果(1)在HEK293A细胞中观察到红色荧光,表明慢病毒转染成功。(2)外源ISCA1蛋白与内源CRY1或CRY2蛋白不结合。(3)与加磁前相比,加磁后Magneto 2.0组细胞的绿色荧光强度增加(1.8±0.5)倍(P<0.05),即发生显著的钙内流;而ISCA1组及细胞对照组细胞的绿色荧光强度与加磁前相比无显著差异。结论外源高表达ISCA1的细胞在此磁场条件刺激后未引起细胞钙内流,无明显磁感应性。OBJECTIVE To explore the effect of iron sulfur cluster binding protein 1(ISCA1)mag⁃netic field on calcium inflow.METHODS①A plasmid containing the ISCA1、Magneto 2.0 gene sequence,and an empty plasmid was prepared for lentivirus packaging,with all the three plasmids carrying the mCherry gene.The infection efficiency of lentiviruses was assessed using fluorescence microscopy.②The immunoprecipitation technique was employed to ascertain the interactions between ISCA1 and cryptochrome(CRY1)as well as CRY2 proteins.③The technique of live cell calcium imaging was used to detect the intracellular calcium influx in cells exhibiting high levels of ISCA1 or Magneto 2.0 expression under the influence of a magnetic field(40 mT,0.1Hz,90%duty cycle).RESULTS①Red fluorescence was observed by fluorescence microscopy,indicating successful transfection of lentivirus.②The exogenous ISCA1 proteins exhibited no affinity towards the endogenous CRY1 or CRY2 proteins.③The green fluorescence intensity of the Magneto 2.0 group increased by(1.8±0.5)times compared to the pre-magnetic addition level,indicating a significant influx of calcium ions.However,there was no significant difference in the green fluorescence intensity between the ISCA1 group and control group before and after magnetic addition.CONCLUSION Cells with exogenously high expres⁃sion of ISCA1 do not elicit calcium influx and exhibit no discernible magnetic induction following stimula⁃tion by this magnetic field.

关 键 词：铁硫簇结合蛋白1 隐花色素 磁受体蛋白 磁感应 磁遗传学 

分 类 号：R96[医药卫生—药理学]