1800MHz射频电磁场通过瞬时受体电位C通道影响原代大鼠皮层神经元突起生长  

Effect of 1800 MHz radiofrequency electromagnetic field on neurite growth of primarily cultured rat cortical neurons via TRPC channel

作　　者：孙立庚 陈纯海[1] 卢永辉[1] 刘川[1] 何旻蒂[1] 高鹏[1] 马秦龙[1] 段维霞[1] 李敏[1] 张蕾[1] 周舟[1] 余争平[1] 钟敏[1] 

机构地区：[1]第三军医大学军事预防医学院劳动卫生学教研室,电磁辐射医学防护教育部重点实验室,重庆400038

出　　处：《第三军医大学学报》2015年第8期772-777,共6页Journal of Third Military Medical University

基　　金：国家自然科学基金面上项目(31270890)~~

摘　　要：目的研究1 800 MHz射频电磁场(radio frequency electromagnetic field,RF-EMF)对原代培养皮层神经元突起生长的影响,并通过瞬时受体电位C通道介导的钙内流,探讨RF-EMF对突起生长影响的机制。方法将原代培养皮层神经元(primarily cultured cortical neurons,PCCNs)用随机数表法分为4组:假暴露(Sham)、Sham+SKF-96365、辐照组(RF)和RF+SKF-96365组。使用s Xc-1800辐照系统,1 800 MHz频率talk模式,5 min开10 min关,比吸收率为4 W/kg,辐照组辐照时间分别为12、24 h和48 h。SKF-96365干预处理是在维持培养基中加入SKF-96365(5μmol/L)后与假暴露组、RF组进行相同的处理。辐照结束后即刻进行以下指标检测:1细胞活力(CCK-8法)和LDH释放;2TRPC1、TRPC3蛋白表达(Western blot);3钙库操纵性钙内流变化(store-operated calcium entry,SOCE)(激光共聚焦显微镜);4神经元突起总长度、初级突起数和分支数(Tuj-1免疫荧光染色)。结果 RF组细胞活力和LDH释放与相应Sham组相比差异均无统计学意义(P>0.05)。与Sham组比较,RF组辐照24 h后,TRPC1和TRPC3蛋白表达分别降低24%和23%,SOCE下降54%,辐照48 h引起PCCNs突起总长度、初级突起数和分支数分别下降20%、12%、31%,以上变化均有统计学意义(P<0.05)。Sham+SKF-96365组SOCE下降24%,突起总长度、初级突起数、分支数分别下降10%、12%、24%,与Sham组比较均有统计学意义(P<0.05),而RF+SKF-96365组SOCE和突起总长度、突起数、分支数的变化与RF组比较差异无统计学意义(P>0.05)。结论 RF-EMF可引起原代皮层神经元突起长度、初级突起数和分支数减少,该效应可能与TRPC1和TRPC3通道蛋白表达降低和TRPC通道介导的钙信号减弱有关。Objective To determine the effect of 1 800 MHz radiofrequency electromagnetic field（ RF-EMF） on neurite growth of primarily cultured cortical neurons（ PCCNs）,and discuss its possible mechanism via transient receptor potential canonical（ TRPC） channel. Methods PCCNs were randomly divided into 4 groups： sham group（ without RF）,sham ＋ SKF-96365（ TRPC blocker） group,RF group,and RF ＋ SKF-96365 group. PCCNs were exposed to RF continuously（ 5 min on and 10 min off） for 12,24 and48 h under 1 800 MHz frequency and talk model,and the specific absorption rate was 4 W / kg. Afterward,the correlation analysis was performed immediately as follows：（ 1） CCK-8 and LDH release,（ 2） Western blot analysis for protein expression of TRPC1 and TRPC3,（ 3） confocal laser scanning microscope-based calcium imaging analysis for store-operated calcium entry（ SOCE）,and（ 4） immunocytochemical analysis for neurite length,neurite number,and neurite branching number. Results The release of CCK-8 and LDH had no statistically significant difference between the RF group and the sham group（ P〈0. 05）. The expression of TRPC1 and TRPC3 proteins was reduced by 24% and 23%,respectively,and SOCE was reduced by 51% in the RF group after 24 h exposure to RF compared with the sham group. After 48 h exposure to RF,the neurite length,neurite number and neurite branching number were reduced by 20%,12% and 31%,respectively,compared with the Sham group,and the differences were significant（ P〈0. 05）. SOCE was reduced by24%,and the neurite length,neurite number and neurite branching number were reduced by 10%,12% and24%,respectively,in the sham ＋ SKF-96365 group compared with the Sham group（ P〈0. 05）. The differences of the SOCE,neurite length,neurite number and neurite branching number between the RF ＋SKF-96365 group and the RF group were not significant（ P〈0. 05）. Conclusion RF-EMF may reduce the neurite length,neurite number,and neurite branching number of the PCCNs,which is related

关 键 词：射频电磁场 瞬时受体电位C 皮层神经元突起 Ca2+ 

分 类 号：R322.81[医药卫生—人体解剖和组织胚胎学] R329.24[医药卫生—基础医学]