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纸质出版:2015
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孙立庚,陈纯海,卢永辉,刘川,何旻蒂,高鹏,马秦龙,段维霞,李敏,张蕾,周舟,余争平,钟敏.1 800 MHz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[J].陆军军医大学学报(原第三军医大学学报),2015,37(08 ):772-777. Sun Ligeng,Chen Chunhai,Lu Yonghui,Liu Chuan,He Mindi,Gao Peng,Ma Qinlong,Duan Wenxia,Li Min,Zhang Lei,Zhou Zhou,Yu Zhengping,Zhong Min.Effect of 1 800 MHz radiofrequency electromagnetic field on neurite growth of primarily cultured rat cortical neurons via TRPC channel[J].J Amry Med Univ (J Third Mil Med Univ),2015,37(08 ):772-777.
孙立庚, 陈纯海, 卢永辉, et al. Effect of 1 800 MHz radiofrequency electromagnetic field on neurite growth of primarily cultured rat cortical neurons via TRPC channel[J]. Journal of Army Medical University, 2015, 37(8): 772-777.
孙立庚,陈纯海,卢永辉,刘川,何旻蒂,高鹏,马秦龙,段维霞,李敏,张蕾,周舟,余争平,钟敏.1 800 MHz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[J].陆军军医大学学报(原第三军医大学学报),2015,37(08 ):772-777. Sun Ligeng,Chen Chunhai,Lu Yonghui,Liu Chuan,He Mindi,Gao Peng,Ma Qinlong,Duan Wenxia,Li Min,Zhang Lei,Zhou Zhou,Yu Zhengping,Zhong Min.Effect of 1 800 MHz radiofrequency electromagnetic field on neurite growth of primarily cultured rat cortical neurons via TRPC channel[J].J Amry Med Univ (J Third Mil Med Univ),2015,37(08 ):772-777. DOI:
孙立庚, 陈纯海, 卢永辉, et al. Effect of 1 800 MHz radiofrequency electromagnetic field on neurite growth of primarily cultured rat cortical neurons via TRPC channel[J]. Journal of Army Medical University, 2015, 37(8): 772-777. DOI:
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sXc-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(5 μmol/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(P0.05)。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(P0.05)。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(P0.05)
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(P0.05)。 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 and 48 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 (P0.05). 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 (P0.05). SOCE was reduced by 24%
and the neurite length
neurite number and neurite branching number were reduced by 10%
12% and 24%
respectively
in the sham+SKF-96365 group compared with the Sham group (P0.05). 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 (P0.05). Conclusion RF-EMF may reduce the neurite length
neurite number
and neurite branching number of the PCCNs
which is related to low expression of TRPC1 and TRPC3 proteins and decrease of TRPC channel-mediated Ca
2+
signals.
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