宽频电磁脉冲辐照对上皮细胞凋亡损伤的影响
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摘要
研究宽频电磁脉冲(Wide-band electromagnetic pulse, WEMP)辐照对正常细胞的损伤作用,寻找辐照参数与细胞凋亡之间的作用规律,为探索WEMP辐照损伤机理模型提供实验依据。利用Cell Counting Kit-8检测WEMP辐照后细胞的活力变化,采用流式细胞术分析细胞凋亡损伤情况,利用统计分析和数据拟合的方法探讨WEMP场强、脉冲个数与细胞凋亡之间的作用规律。结果表明,在本实验条件下,场强与脉冲个数都是引起人羊膜上皮细胞(AECs)凋亡率增加的因素,其中,电场强度为主要因素,脉冲个数为次要因素,两者之间存在交互作用。细胞凋亡率与电场强度之间呈现出Logistic函数规律。
The purpose of this study was to investigate the relationship between cell apoptosis and wide-band electromagnetic pulse(WEMP) irradiation parameters and to provide experimental data to establish the biophysics damage model for WEMP. Cell counting kit-8 was used to detect the cell viability changes after WEMP irradiation and flow cytometry assay was used to detect cell apoptosis after irradiation under different WEMP irradiation parameters. Under experimental conditions, field strength and number of pulses were observed to be the main factors that increased the apoptotic rate of human amniotic epithelial cells. Specifically, electric field intensity and number of pulses were determined to be the primary and secondary factors, respectively. The two factors were confirmed to interact with each other, that is, a logistic function was found to exist between the apoptotic rate and electric field strength.
The purpose of this study was to investigate the relationship between cell apoptosis and wide-band electromagnetic pulse(WEMP) irradiation parameters and to provide experimental data to establish the biophysics damage model for WEMP. Cell counting kit-8 was used to detect the cell viability changes after WEMP irradiation and flow cytometry assay was used to detect cell apoptosis after irradiation under different WEMP irradiation parameters. Under experimental conditions, field strength and number of pulses were observed to be the main factors that increased the apoptotic rate of human amniotic epithelial cells. Specifically, electric field intensity and number of pulses were determined to be the primary and secondary factors, respectively. The two factors were confirmed to interact with each other, that is, a logistic function was found to exist between the apoptotic rate and electric field strength.
引文
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4李玉红,王德文,王水明,等.电磁脉冲辐射后海马形态学观察及凋亡相关基因检测[J].解放军预防医学杂志2006,24(2):95-98.DOI:10.13704/j.cnki.jyyx.2006.02.00LI Yuhong,WANG Dewen,WANG Shuiming Morphological changes of hippocampus and the expression of apoptosis-associated genes in rats exposed to EMP[J].Medical Journal of Chinese People’s Liberation Army,2006,24(2):95-98.DOI:10.13704/j cnki.jyyx.2006.02.00.
5张俊平,郭玲,张克英,等.射频辐射诱导小鼠焦虑样行为的机制探讨[J].神经解剖学杂志,2018,34(5):541-547.DOI:10.16557/j.cnki.10007547.2018.05.003.ZHANG Junping,GUO Ling,ZHANG Keying,et al Study on the mechanisms of radiofrequency radiation induced anxiety-like behavior in mice[J].Chinese Journal of Neuroanatomy,2018,34(5):541-547.DOI:10.16557/j cnki.10007547.2018.05.003.
6杨孔,秦天莺,岳碧松,等.电磁脉冲对肿瘤细胞的影响[J].四川大学学报,2006,43(4):894-898.DOI:10.1016/S1872-2075(06)60069-1.YANG Kong,QIN Tianying,YUE Bisong,et al.Effects of electromagnetic pulses on tumor cells[J].Journal of Sichuan University,2006,43(4):894-898.DOI:10.1016/S1872-2075(06)60069-1.
7 Ibey B L,Xiao S,Schoenbach K H,et al.Plasma membrane permeabilization by 60-and 600-ns electric pulses is determined by the absorbed dose[J]Bioelectromagnetics,2009,30(2):92-99.DOI:10.1002/bem.20451.
8 Fini M,Giavaresi G,Carpi A,et al.Effects of pulsed electromagnetic fields on articular hyaline cartilage:a review of experimental and clinical studies[J]Biomedicine&Pharmacotherapy,2005,59(7):388-394DOI:10.1016/j.biopha.2005.02.002.
9 Kim Y M,Kim H S,Lee J S,et al.Effects of 60 Hz 14microT magnetic field on the apoptosis of testicular germ cell in mice[J].Bioelecteomagnetics,2009,30(1):66-72DOI:10.1002/bem.20448.
10 Kumar S,Behari J,Sisodia R.Influence of electromagnetic fields on reproductive system of male rats[J].International Journal of Radiation Biology,201389(3):147-154.DOI:10.3109/09553002.2013.741282.
11罗亚宁,丁桂荣,陈永斌,等.电磁脉冲辐照对小鼠睾丸组织凋亡和转化生长因子β3表达的影响[J].中华劳动卫生职业病杂志,2014,32(4):251-255.DOI:10.3760cma.j.issn.1001-9391.2014.04.003.LUO Yaning,DING Guirong,CHEN Yongbin,et al Effects of electromagnetic pulses on apoptosis and TGF-p3 expression of mouse testis tissue[J].Chinese Journa of Industrial Hygiene and Occupational Diseases,201432(4):251-255.DOI:10.3760/cma.j.issn.1001-93912014.04.003.
12林艳云,王亚峰,高鹏,等.电磁脉冲暴露对幼龄雄性小鼠睾丸发育的影响[J].辐射研究与辐射工艺学报2018,36(4):040202.DOI:10.11889/j.1000-3436.2018rrj.36.040202.LIN Yanyun,WANG Yafeng,GAO Peng,et al.Effects of electromagnetic pluse exposure on development of testicle in the growing male mice[J].Journal of Radiation Research and Radiation Processing,2018,36(4):040202DOI:10.11889/j.1000-3436.2018.rrj.36.040202.
13崔玉芳,杨红,高亚兵,等.中低强度电磁脉冲诱发大鼠淋巴细胞凋亡及其机制的研究[J].感染、炎症、修复2004,5(1,2):19-21.CUI Yufang,YANG Hong,GAO Yabing,et al.Apoptosis of lymphocytes induced by low and moderate dose of electromagnetic pulse and its mechanism[J].Infection Inflammation,Repair,2004,5(1,2):19-21.
14王谦,吴文超,陈晓莹,等.不同频率脉冲电磁场对人卵巢癌SKOV3细胞增殖、凋亡及迁移的影响[J].生物医学工程学杂志,2012,29(2):291-295.WANG Qian,WU Wenchao,CHEN Xiaoying,et al Effect of pulsed electromagnetic field with different frequencies on the proliferation,apoptosis and migration of human ovarian cancer cells[J].Journal of Biomedical Engineering,2012,29(2):291-295.
15白孟海,葛宝丰,韦哲,等.脉冲电磁场对去势大鼠骨髓源细胞体外培养破骨细胞变化的影响[J].中国骨伤2009,22(10):727-729.BAI Menghai,GE Baofeng,WEI Zhe,et al.Effect of pulsed electromagnetic field on the changes of osteoclasts in ovariectomized rats bone marrow culture in vitro[J]China Journal of Orthopaedics and Traumatology,200922(10):727-729.
16 Wang X H,Qi H X,Zhang J,et al.Multivariable quantitative relation between cell viability and the exposure parameters of 9.33 GHz RF-EMP irradiation[J]Electromagnetic Biology and Medicine,2018,37(3):146-154.DOI:10.1080/15368378.2018.1482221.
17刘合,雷鸣嘉,张杰,等.宽频电磁脉冲辐照引起大鼠生殖系统的损伤与修复[J].辐射研究与辐射工艺学报2017,35(1):010203.DOI:10.11889/j.1000-3436.2017rrj.35.010203.LIU He,LEI Mingjia,ZHANG Jie,et al.Effect of wideband electromagnetic pulses on the reproductive system of rat and subsequent recovery[J].Journal of Radiation Research and Radiation Processing,2017,35(1):010203DOI:10.11889/j.1000-3436.2017.rrj.35.010203.
18林锦文,刘合,张杰,等.电磁脉冲对大鼠心脏和肝脏损伤及修复的影响[J].辐射研究与辐射工艺学报,201533(6):060201.DOI:10.11889/j.1000-3436.2015.rrj.33060201.LIN Jinwen,LIU He,ZHANG Jie,et al.Effects of electromagnetic pulses on the heart and liver of rat and subsequent self-recovery[J].Journal of Radiation Research and Radiation Processing,2015,33(6):060201DOI:10.11889/j.1000-3436.2015.rrj.33.060201.
19雷鸣嘉,刘合,张杰,等.宽频电磁脉冲多次辐照对雄性大鼠生殖系统的累积损伤与修复[J].辐射研究与辐射工艺学报,2018,36(1):010202.DOI:10.11889/j.1000-3436.2018.rrj.36.010202.LEI Mingjia,LIU He,ZHANG Jie,et al.Cumulative effect of wide-band electromagnetic pulse exposure on the reproductive system of male rats[J].Journal of Radiation Research and Radiation Processing,2018,36(1):010202.DOI:10.11889/j.1000-3436.2018rrj.36.010202.
20李好山,熊正爱,姚陈果.脉冲电场诱导细胞凋亡机制及其在肿瘤治疗中应用前景展望[J].重庆医学,201140(20):2059-2061.DOI:10.3969/j.issn.1671-83482011.20.041.LI Haoshan,XIONG Zhengai,YAO Chenguo Mechanism of pulsed electric field induced apoptosis and its application prospect in tumor therapy[J].Chongqing Medical Journal,2011,40(20):2059-2061.DOI:10.3969j.issn.1671-8348.2011.20.041.
21侯书胜,王书平.核电磁脉冲模拟装置[J].高电压技术2002,28(1):35-36+57.HOU Shusheng,WANG Shuping.Simulation of nuclear electromagnetic pulse[J].High Voltage Engineering2002,28(1):35-36+57.
22邱志斌,阮江军,徐闻婕,等.典型电极短空气间隙的击穿电压混合预测[J].高电压技术，2018,44(6):2012-2018.DOI:10.13336/j.1003-6520.hve.20180529038.QIU Zhibin,RUAN Jiangjun,XU Wenjie,et al.Hybrid prediction of the breakdown voltages of short air gaps with typical electrodes[J].High Voltage Engineering2018,44(6):2012-2018.DOI:10.13336/j.1003-6520hve.20180529038.
2王甜,郭玲,安广洲,等.1950 MHz射频电磁场对人脐带间充质干细胞增殖和成骨分化的影响研究[J].现代生物医学进展,2018,18(12):2218-2223.DOI:10.13241j.cnki.pmb.2018.12.004.WANG Tian,GUO Ling,AN Guangzhou,et al.Effects of 1950 MHz radiofrequency electromagnetic fields on the proliferation and osteogenic differentiation of human umbilical cord mesenchymal stem cells[J].Progress in Modern Biomedicine,2018,18(12):2218-2223.DOI10.13241/j.cnki.pmb.2018.12.004.
3 Li K C,Ma S R,Ren D Q,et al.Effects of electromagnetic pulse on serum element levels in rat[J]Biological Trace Element Research,2014,158(1):81-86DOI:10.1007/s12011-014-9903-0.
4李玉红,王德文,王水明,等.电磁脉冲辐射后海马形态学观察及凋亡相关基因检测[J].解放军预防医学杂志2006,24(2):95-98.DOI:10.13704/j.cnki.jyyx.2006.02.00LI Yuhong,WANG Dewen,WANG Shuiming Morphological changes of hippocampus and the expression of apoptosis-associated genes in rats exposed to EMP[J].Medical Journal of Chinese People’s Liberation Army,2006,24(2):95-98.DOI:10.13704/j cnki.jyyx.2006.02.00.
5张俊平,郭玲,张克英,等.射频辐射诱导小鼠焦虑样行为的机制探讨[J].神经解剖学杂志,2018,34(5):541-547.DOI:10.16557/j.cnki.10007547.2018.05.003.ZHANG Junping,GUO Ling,ZHANG Keying,et al Study on the mechanisms of radiofrequency radiation induced anxiety-like behavior in mice[J].Chinese Journal of Neuroanatomy,2018,34(5):541-547.DOI:10.16557/j cnki.10007547.2018.05.003.
6杨孔,秦天莺,岳碧松,等.电磁脉冲对肿瘤细胞的影响[J].四川大学学报,2006,43(4):894-898.DOI:10.1016/S1872-2075(06)60069-1.YANG Kong,QIN Tianying,YUE Bisong,et al.Effects of electromagnetic pulses on tumor cells[J].Journal of Sichuan University,2006,43(4):894-898.DOI:10.1016/S1872-2075(06)60069-1.
7 Ibey B L,Xiao S,Schoenbach K H,et al.Plasma membrane permeabilization by 60-and 600-ns electric pulses is determined by the absorbed dose[J]Bioelectromagnetics,2009,30(2):92-99.DOI:10.1002/bem.20451.
8 Fini M,Giavaresi G,Carpi A,et al.Effects of pulsed electromagnetic fields on articular hyaline cartilage:a review of experimental and clinical studies[J]Biomedicine&Pharmacotherapy,2005,59(7):388-394DOI:10.1016/j.biopha.2005.02.002.
9 Kim Y M,Kim H S,Lee J S,et al.Effects of 60 Hz 14microT magnetic field on the apoptosis of testicular germ cell in mice[J].Bioelecteomagnetics,2009,30(1):66-72DOI:10.1002/bem.20448.
10 Kumar S,Behari J,Sisodia R.Influence of electromagnetic fields on reproductive system of male rats[J].International Journal of Radiation Biology,201389(3):147-154.DOI:10.3109/09553002.2013.741282.
11罗亚宁,丁桂荣,陈永斌,等.电磁脉冲辐照对小鼠睾丸组织凋亡和转化生长因子β3表达的影响[J].中华劳动卫生职业病杂志,2014,32(4):251-255.DOI:10.3760cma.j.issn.1001-9391.2014.04.003.LUO Yaning,DING Guirong,CHEN Yongbin,et al Effects of electromagnetic pulses on apoptosis and TGF-p3 expression of mouse testis tissue[J].Chinese Journa of Industrial Hygiene and Occupational Diseases,201432(4):251-255.DOI:10.3760/cma.j.issn.1001-93912014.04.003.
12林艳云,王亚峰,高鹏,等.电磁脉冲暴露对幼龄雄性小鼠睾丸发育的影响[J].辐射研究与辐射工艺学报2018,36(4):040202.DOI:10.11889/j.1000-3436.2018rrj.36.040202.LIN Yanyun,WANG Yafeng,GAO Peng,et al.Effects of electromagnetic pluse exposure on development of testicle in the growing male mice[J].Journal of Radiation Research and Radiation Processing,2018,36(4):040202DOI:10.11889/j.1000-3436.2018.rrj.36.040202.
13崔玉芳,杨红,高亚兵,等.中低强度电磁脉冲诱发大鼠淋巴细胞凋亡及其机制的研究[J].感染、炎症、修复2004,5(1,2):19-21.CUI Yufang,YANG Hong,GAO Yabing,et al.Apoptosis of lymphocytes induced by low and moderate dose of electromagnetic pulse and its mechanism[J].Infection Inflammation,Repair,2004,5(1,2):19-21.
14王谦,吴文超,陈晓莹,等.不同频率脉冲电磁场对人卵巢癌SKOV3细胞增殖、凋亡及迁移的影响[J].生物医学工程学杂志,2012,29(2):291-295.WANG Qian,WU Wenchao,CHEN Xiaoying,et al Effect of pulsed electromagnetic field with different frequencies on the proliferation,apoptosis and migration of human ovarian cancer cells[J].Journal of Biomedical Engineering,2012,29(2):291-295.
15白孟海,葛宝丰,韦哲,等.脉冲电磁场对去势大鼠骨髓源细胞体外培养破骨细胞变化的影响[J].中国骨伤2009,22(10):727-729.BAI Menghai,GE Baofeng,WEI Zhe,et al.Effect of pulsed electromagnetic field on the changes of osteoclasts in ovariectomized rats bone marrow culture in vitro[J]China Journal of Orthopaedics and Traumatology,200922(10):727-729.
16 Wang X H,Qi H X,Zhang J,et al.Multivariable quantitative relation between cell viability and the exposure parameters of 9.33 GHz RF-EMP irradiation[J]Electromagnetic Biology and Medicine,2018,37(3):146-154.DOI:10.1080/15368378.2018.1482221.
17刘合,雷鸣嘉,张杰,等.宽频电磁脉冲辐照引起大鼠生殖系统的损伤与修复[J].辐射研究与辐射工艺学报2017,35(1):010203.DOI:10.11889/j.1000-3436.2017rrj.35.010203.LIU He,LEI Mingjia,ZHANG Jie,et al.Effect of wideband electromagnetic pulses on the reproductive system of rat and subsequent recovery[J].Journal of Radiation Research and Radiation Processing,2017,35(1):010203DOI:10.11889/j.1000-3436.2017.rrj.35.010203.
18林锦文,刘合,张杰,等.电磁脉冲对大鼠心脏和肝脏损伤及修复的影响[J].辐射研究与辐射工艺学报,201533(6):060201.DOI:10.11889/j.1000-3436.2015.rrj.33060201.LIN Jinwen,LIU He,ZHANG Jie,et al.Effects of electromagnetic pulses on the heart and liver of rat and subsequent self-recovery[J].Journal of Radiation Research and Radiation Processing,2015,33(6):060201DOI:10.11889/j.1000-3436.2015.rrj.33.060201.
19雷鸣嘉,刘合,张杰,等.宽频电磁脉冲多次辐照对雄性大鼠生殖系统的累积损伤与修复[J].辐射研究与辐射工艺学报,2018,36(1):010202.DOI:10.11889/j.1000-3436.2018.rrj.36.010202.LEI Mingjia,LIU He,ZHANG Jie,et al.Cumulative effect of wide-band electromagnetic pulse exposure on the reproductive system of male rats[J].Journal of Radiation Research and Radiation Processing,2018,36(1):010202.DOI:10.11889/j.1000-3436.2018rrj.36.010202.
20李好山,熊正爱,姚陈果.脉冲电场诱导细胞凋亡机制及其在肿瘤治疗中应用前景展望[J].重庆医学,201140(20):2059-2061.DOI:10.3969/j.issn.1671-83482011.20.041.LI Haoshan,XIONG Zhengai,YAO Chenguo Mechanism of pulsed electric field induced apoptosis and its application prospect in tumor therapy[J].Chongqing Medical Journal,2011,40(20):2059-2061.DOI:10.3969j.issn.1671-8348.2011.20.041.
21侯书胜,王书平.核电磁脉冲模拟装置[J].高电压技术2002,28(1):35-36+57.HOU Shusheng,WANG Shuping.Simulation of nuclear electromagnetic pulse[J].High Voltage Engineering2002,28(1):35-36+57.
22邱志斌,阮江军,徐闻婕,等.典型电极短空气间隙的击穿电压混合预测[J].高电压技术，2018,44(6):2012-2018.DOI:10.13336/j.1003-6520.hve.20180529038.QIU Zhibin,RUAN Jiangjun,XU Wenjie,et al.Hybrid prediction of the breakdown voltages of short air gaps with typical electrodes[J].High Voltage Engineering2018,44(6):2012-2018.DOI:10.13336/j.1003-6520hve.20180529038.
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