1.8 GHz射频电磁辐射对人晶状体上皮细胞氧化应激影响的研究
发布时间：2018-10-29 14:30
【摘要】：背景 随着的移动通讯设备的广泛使用,射频电磁辐射可能带来的不良效应越来越受到大众的关注。氧化应激系体内氧化与抗氧化失衡从而导致了机体生理生化改变和多种慢性疾病,因此射频电磁辐射与氧化应激的关系近年来已成为人们的研究方向。然而低强度射频电磁辐射能否诱导氧化应激目前还存在争议,且射频电磁辐射诱导氧化应激的机制也不清楚。已知很多眼科疾病和氧化应激有关,射频电磁辐射诱发晶状体上皮细胞的生物学效应也有大量报道。因此,研究低强度射频电磁辐射是否能诱导晶状体上皮细胞氧化应激,并且对诱导氧化应激的机理做进一步研究,将有助于了解射频电磁辐射所致晶状体上皮细胞损伤的机制。 目的 探讨1.8GHz射频电磁辐射对体外培养的人晶状体上皮细胞氧化应激的影响以及可能的机制。 方法 体外培养人晶状体上皮细胞(hLECs)分为辐照组与假辐照组。同时将其置于1.8GHz射频电磁场内进行间断(5分钟开,10分钟停)辐照,辐照组辐照强度比吸收率(specific absorption rate, SAR)为2,3,或4W/kg,假辐照为0W/kg。经过0.5,1,或1.5小时辐照后,立刻使用荧光探针2’,7’-二氯荧光素二乙酸酯(2',7'-dichlorofluorescin diacetate, DCFH-DA)检测细胞内活性氧(reactive oxygen species, ROS)水平。经过6,12,或24小时辐照后,使用丙二醛(malondialdehyde, MDA)检测试剂盒测量细胞脂质过氧化水平。经过6,12,或24小时辐照后,使用CCK-8(Cell Counting Kit-8)试剂盒检测细胞活力。 运用实时定量PCR技术在转录水平研究1.8GHz射频电磁辐射对抗氧化基因SOD1, SOD2, CAT和GPX1表达的影响。人晶状体上皮细胞接受1小时辐照,提取总RNA经逆转录后用实时荧光定量PCR试验检测上述4个抗氧化基因的表达水平。 运用Western blot技术在蛋白水平研究1.8GHz射频电磁辐射对抗氧化酶SOD1, SOD2, CAT和GPx1的影响。人晶状体上皮细胞接受1小时辐照,提取总蛋白后用使用Western blot检测上述4个抗氧化酶蛋白的表达水平。 结果 经过0.5,1,或1.5小时1.8GHz射频电磁场辐照后,辐照组晶状体上皮细胞内的ROS水平明显高于假辐照组(P0.05)。经过6,12,或24小时辐照后,辐照组细胞培养基中的MDA浓度水平明显高于假辐照组(P0.05)。经过6,12,或24小时辐照后,辐照组细胞细胞活力明显低于假辐照组(P0.05)。实时定量RT-PCR结果显示经过1小时辐照,辐照组细胞SOD1, SOD2, CAT和GPX1的mRNA表达显著低于假辐照组(P0.05)。Western blot试验显示经过1小时辐照,辐照组细胞SOD1, SOD2,CAT和GPx1蛋白表达明显低于假辐照组(P0.05)。 结论 低强度1.8GHz射频电磁辐射能使得晶状体上皮细胞产生氧化应激。细胞内ROS水平上升可能和射频电磁辐射下抗氧化酶基因下调有关。
[Abstract]:Background with the wide use of mobile communication devices, the potential adverse effects of RF electromagnetic radiation have attracted more and more attention. Oxidative stress is the imbalance between oxidation and oxidation in vivo, which leads to physiological and biochemical changes and many chronic diseases. Therefore, the relationship between radiofrequency electromagnetic radiation and oxidative stress has become a research direction in recent years. However, whether low intensity radiofrequency electromagnetic radiation can induce oxidative stress is still controversial, and the mechanism of radiofrequency electromagnetic radiation induced oxidative stress is not clear. Many eye diseases are known to be related to oxidative stress, and the biological effects of radiofrequency electromagnetic radiation on lens epithelial cells have also been reported. Therefore, to study whether low intensity radiofrequency electromagnetic radiation can induce oxidative stress of lens epithelial cells, and to further study the mechanism of inducing oxidative stress, will be helpful to understand the mechanism of lens epithelial cell injury induced by radio frequency electromagnetic radiation. Objective to investigate the effect of 1.8GHz radiofrequency electromagnetic radiation on oxidative stress in cultured human lens epithelial cells and its possible mechanism. Methods cultured human lens epithelial cells (hLECs) were divided into irradiation group and false irradiation group. At the same time, it was placed in the 1.8GHz radio frequency electromagnetic field for intermittent irradiation (5 minutes open, 10 minutes stop). The radiation intensity of the irradiation group was 2W / kg, or 4W / kg, and the radiation intensity was 0 W / kg. The specific absorptivity of the irradiation group was 2W / kg, or 4W / kg. After 0. 5 or 1. 5 hours of irradiation, the level of reactive oxygen species (Ros) (reactive oxygen species, ROS) in the cells was detected by fluorescence probe 2H 7G-dichlorofluorescin diacetate, DCFH-DA. The lipid peroxidation level was measured by malondialdehyde (MDA) (malondialdehyde, MDA) assay kit after irradiation for 6 ~ 12, or 24 hours. Cell viability was detected by CCK-8 (Cell Counting Kit-8 kit after irradiation for 6 ~ 12, or 24 hours. The effect of 1.8GHz radiofrequency electromagnetic radiation on the expression of antioxidant genes SOD1, SOD2, CAT and GPX1 was studied at transcriptional level by real-time quantitative PCR. Human lens epithelial cells (LECs) were irradiated for 1 hour. The total RNA was extracted by reverse transcription and the expression levels of the above four antioxidant genes were detected by real-time fluorescence quantitative PCR assay. The effects of 1.8GHz RF electromagnetic radiation on SOD1, SOD2, CAT and GPx1 were studied at protein level by Western blot technique. Human lens epithelial cells were irradiated for 1 hour. The total protein was extracted and the expression of the above four antioxidant enzyme proteins was detected by Western blot. Results the ROS level of lens epithelial cells in the irradiated group was significantly higher than that in the sham irradiation group (P0.05). The MDA concentration in the cell culture medium of the irradiated group was significantly higher than that of the sham irradiation group (P0.05). The cell viability in the irradiation group was significantly lower than that in the sham irradiation group (P0.05). The results of real-time quantitative RT-PCR showed that after 1 hour irradiation, the mRNA expression of SOD1, SOD2, CAT and GPX1 in irradiated cells was significantly lower than that in false irradiation group (P0.05). Western blot test showed that after 1 hour irradiation, the SOD1, SOD2, expression of irradiated cells was significantly lower than that of false irradiation group). The expression of CAT and GPx1 protein was significantly lower than that of false irradiation group (P0.05). Conclusion low intensity 1.8GHz radiofrequency electromagnetic radiation can induce oxidative stress in lens epithelial cells. The increase of intracellular ROS level may be related to the down-regulation of antioxidant enzyme gene under radiofrequency electromagnetic radiation.
【学位授予单位】：浙江大学
【学位级别】：博士
【学位授予年份】：2013
【分类号】：R779.1
[Abstract]:Background with the wide use of mobile communication devices, the potential adverse effects of RF electromagnetic radiation have attracted more and more attention. Oxidative stress is the imbalance between oxidation and oxidation in vivo, which leads to physiological and biochemical changes and many chronic diseases. Therefore, the relationship between radiofrequency electromagnetic radiation and oxidative stress has become a research direction in recent years. However, whether low intensity radiofrequency electromagnetic radiation can induce oxidative stress is still controversial, and the mechanism of radiofrequency electromagnetic radiation induced oxidative stress is not clear. Many eye diseases are known to be related to oxidative stress, and the biological effects of radiofrequency electromagnetic radiation on lens epithelial cells have also been reported. Therefore, to study whether low intensity radiofrequency electromagnetic radiation can induce oxidative stress of lens epithelial cells, and to further study the mechanism of inducing oxidative stress, will be helpful to understand the mechanism of lens epithelial cell injury induced by radio frequency electromagnetic radiation. Objective to investigate the effect of 1.8GHz radiofrequency electromagnetic radiation on oxidative stress in cultured human lens epithelial cells and its possible mechanism. Methods cultured human lens epithelial cells (hLECs) were divided into irradiation group and false irradiation group. At the same time, it was placed in the 1.8GHz radio frequency electromagnetic field for intermittent irradiation (5 minutes open, 10 minutes stop). The radiation intensity of the irradiation group was 2W / kg, or 4W / kg, and the radiation intensity was 0 W / kg. The specific absorptivity of the irradiation group was 2W / kg, or 4W / kg. After 0. 5 or 1. 5 hours of irradiation, the level of reactive oxygen species (Ros) (reactive oxygen species, ROS) in the cells was detected by fluorescence probe 2H 7G-dichlorofluorescin diacetate, DCFH-DA. The lipid peroxidation level was measured by malondialdehyde (MDA) (malondialdehyde, MDA) assay kit after irradiation for 6 ~ 12, or 24 hours. Cell viability was detected by CCK-8 (Cell Counting Kit-8 kit after irradiation for 6 ~ 12, or 24 hours. The effect of 1.8GHz radiofrequency electromagnetic radiation on the expression of antioxidant genes SOD1, SOD2, CAT and GPX1 was studied at transcriptional level by real-time quantitative PCR. Human lens epithelial cells (LECs) were irradiated for 1 hour. The total RNA was extracted by reverse transcription and the expression levels of the above four antioxidant genes were detected by real-time fluorescence quantitative PCR assay. The effects of 1.8GHz RF electromagnetic radiation on SOD1, SOD2, CAT and GPx1 were studied at protein level by Western blot technique. Human lens epithelial cells were irradiated for 1 hour. The total protein was extracted and the expression of the above four antioxidant enzyme proteins was detected by Western blot. Results the ROS level of lens epithelial cells in the irradiated group was significantly higher than that in the sham irradiation group (P0.05). The MDA concentration in the cell culture medium of the irradiated group was significantly higher than that of the sham irradiation group (P0.05). The cell viability in the irradiation group was significantly lower than that in the sham irradiation group (P0.05). The results of real-time quantitative RT-PCR showed that after 1 hour irradiation, the mRNA expression of SOD1, SOD2, CAT and GPX1 in irradiated cells was significantly lower than that in false irradiation group (P0.05). Western blot test showed that after 1 hour irradiation, the SOD1, SOD2, expression of irradiated cells was significantly lower than that of false irradiation group). The expression of CAT and GPx1 protein was significantly lower than that of false irradiation group (P0.05). Conclusion low intensity 1.8GHz radiofrequency electromagnetic radiation can induce oxidative stress in lens epithelial cells. The increase of intracellular ROS level may be related to the down-regulation of antioxidant enzyme gene under radiofrequency electromagnetic radiation.
【学位授予单位】：浙江大学
【学位级别】：博士
【学位授予年份】：2013
【分类号】：R779.1
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