Zeng Qunli, Hu Genlin, Chiang Huai, Fu Yiti, Mao Guogen, Lu Deqiang
Microwave Lab., Zhejiang University School of Medicine, Hangzhou 310031, China.
Zhonghua Lao Dong Wei Sheng Zhi Ye Bing Za Zhi. 2002 Aug;20(4):260-2.
To study the effects of extremely low frequency magnetic fields(ELF MF) on the amount and localization of connexin 43(Cx43) gap-junction protein in the Chinese hamster lung(CHL) cells, and to explore the mechanism of ELF MF suppression on gap-junctional intercellular communication(GJIC).
The cells were irradiated for 24 h with 50 Hz sinusoidal magnetic field at 0.8 mT without or with 12-O-tetrade-canoylphorbol-3-acetate(TPA), 5 ng/ml for 1 h. The localization of Cx43 proteins were performed by indirect immunofluorescence histochemical analysis and detected by confocal microscopy. The second experiment was conducted to examine the quantity of Cx43 proteins level in nuclei or cytoplasm and detected by Western blotting analysis.
The cells exposed to TPA for 1 h displayed less bright labelled spots in the regions of intercellular junction than the normal cells. Most of Cx43 labelled spots occurred in the cytoplasm and aggregated near the nuclei. At the same time, the amount of Cx43 protein in cytoplasm were increased[(2.03 +/- 0.89) in ELF group, (2.43 +/- 0.82) in TPA group] as compared to normal control(1.04 +/- 0.17) (P < 0.01).
Inhibition on GJIC function by ELF MF alone or combined with TPA may be related with the shift of Cx43 from the regions of intercellular junction to the cytoplasm.
研究极低频磁场(ELF MF)对中国仓鼠肺(CHL)细胞中连接蛋白43(Cx43)间隙连接蛋白数量和定位的影响,并探讨ELF MF抑制间隙连接细胞间通讯(GJIC)的机制。
将细胞暴露于0.8 mT的50 Hz正弦磁场中24小时,有无12 - O - 十四烷酰佛波醇 - 13 - 乙酸酯(TPA)(5 ng/ml,作用1小时)。通过间接免疫荧光组织化学分析进行Cx43蛋白的定位,并用共聚焦显微镜检测。进行第二个实验以检测细胞核或细胞质中Cx43蛋白水平的数量,并用蛋白质免疫印迹分析进行检测。
暴露于TPA 1小时的细胞在细胞间连接区域显示出比正常细胞更少的明亮标记斑点。大多数Cx43标记斑点出现在细胞质中并聚集在细胞核附近。同时,与正常对照(1.04±0.17)相比,细胞质中Cx43蛋白的量增加[ELF组为(2.03±0.89),TPA组为(2.43±0.82)](P < 0.01)。
ELF MF单独或与TPA联合对GJIC功能的抑制可能与Cx从细胞间连接区域向细胞质的转移有关。