应用γ-H2AX分析技术评价工频磁场对不同细胞DNA双链断裂的影响
发布时间：2018-12-07 13:38
【摘要】：目的工频磁场是频率为50Hz的极低频磁场,随着人们接触频率及强度的提高,工频磁场的生物学效应引起了研究者的极大关注,有关工频磁场对细胞DNA损伤修复的影响成为评价其健康风险的关键事件之一,但迄今为止有关其遗传损伤效应的研究报道尚无确切定论。本研究采用DNA双链断裂的早期敏感指标—γH2AX,初步评价了工频磁场不同辐照强度和时间对三种不同系统或器官来源细胞的DNA损伤效应,以期为工频磁场敏感细胞的筛选及其进一步的生物学效应研究奠定基础。 方法将人羊膜上皮细胞(FL)、人脐静脉内皮细胞(HUVEC)以及人皮肤成纤维细胞(HSF)分别暴露于50Hz,0.4mT、1mT和2mT工频磁场15min、1h和24h后,采用γH2AX免疫荧光法和免疫印迹法分别检测了细胞内DNA双链断裂和γH2AX表达情况。每个强度和时间点分别设置一一对应假暴露组,同时采用0.5μM或1μM4NQO处理1h作为阳性对照组。其中,γH2AX免疫荧光实验采用平均焦点数、焦点阳性细胞率以及焦点分段分布率作为DNA损伤程度的评价指标。实验采用双盲,每组实验重复3-5次。 结果γH2AX免疫荧光法和免疫印迹法结果显示,工频磁场各暴露组与一一对应假暴露组相比,平均焦点数、焦点阳性细胞率、yH2AX蛋白表达量均没有显著性差异(P0.05),焦点分段分布率在个别强度或时间点上有差异(P0.05)。 结论在本实验条件下,工频磁场未能导致FL、HUVEC以及HSF细胞DNA损伤。
[Abstract]:Aim Power frequency magnetic field is an extremely low frequency magnetic field with 50Hz frequency. With the increase of contact frequency and intensity, the biological effect of power frequency magnetic field has attracted great attention of researchers. The effect of power frequency magnetic field on the repair of DNA damage has become one of the key events in evaluating the health risk of cells. In this study, the early sensitive index of DNA double strand break, 纬 H 2AX, was used to evaluate the DNA damage effect of different radiation intensity and time of power frequency magnetic field on cells from three different systems or organs. In order to lay a foundation for the screening of power frequency magnetic field sensitive cells and the further study of biological effects. Methods Human amniotic epithelial cells (FL), human umbilical vein endothelial cells (HUVEC) and human skin fibroblasts (HSF) were exposed to 50 Hz 0. 4 mTT 1 Mt and 2mT power frequency magnetic field for 1 h and 24 h, respectively. The double strand breaks of DNA and the expression of 纬 H2AX in the cells were detected by 纬 H2AX immunofluorescence assay and Western blot respectively. Each intensity and time point were treated with 0. 5 渭 M or 1 渭 M4NQO for 1 h as the positive control group. In the 纬 H2AX immunofluorescence assay, the average number of focal points, the percentage of focal positive cells and the distribution rate of focal points were used to evaluate the degree of DNA damage. The experiment was double-blind and repeated 3-5 times in each group. Results the results of 纬 H2AX immunofluorescence assay and Western blotting showed that there was no significant difference in the number of focal points, the percentage of focal positive cells and the expression of yH2AX protein between the power frequency magnetic field exposure group and the one-to-one corresponding false exposure group (P0.05). The distribution rate of focus segment was different in individual intensity or time point (P0.05). Conclusion under this condition, the power frequency magnetic field can not cause DNA damage of FL,HUVEC and HSF cells.
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R142
本文编号：2367287
[Abstract]:Aim Power frequency magnetic field is an extremely low frequency magnetic field with 50Hz frequency. With the increase of contact frequency and intensity, the biological effect of power frequency magnetic field has attracted great attention of researchers. The effect of power frequency magnetic field on the repair of DNA damage has become one of the key events in evaluating the health risk of cells. In this study, the early sensitive index of DNA double strand break, 纬 H 2AX, was used to evaluate the DNA damage effect of different radiation intensity and time of power frequency magnetic field on cells from three different systems or organs. In order to lay a foundation for the screening of power frequency magnetic field sensitive cells and the further study of biological effects. Methods Human amniotic epithelial cells (FL), human umbilical vein endothelial cells (HUVEC) and human skin fibroblasts (HSF) were exposed to 50 Hz 0. 4 mTT 1 Mt and 2mT power frequency magnetic field for 1 h and 24 h, respectively. The double strand breaks of DNA and the expression of 纬 H2AX in the cells were detected by 纬 H2AX immunofluorescence assay and Western blot respectively. Each intensity and time point were treated with 0. 5 渭 M or 1 渭 M4NQO for 1 h as the positive control group. In the 纬 H2AX immunofluorescence assay, the average number of focal points, the percentage of focal positive cells and the distribution rate of focal points were used to evaluate the degree of DNA damage. The experiment was double-blind and repeated 3-5 times in each group. Results the results of 纬 H2AX immunofluorescence assay and Western blotting showed that there was no significant difference in the number of focal points, the percentage of focal positive cells and the expression of yH2AX protein between the power frequency magnetic field exposure group and the one-to-one corresponding false exposure group (P0.05). The distribution rate of focus segment was different in individual intensity or time point (P0.05). Conclusion under this condition, the power frequency magnetic field can not cause DNA damage of FL,HUVEC and HSF cells.
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R142
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