本文選題：射頻電磁場(chǎng) + 相對(duì)敏感細(xì)胞��； 參考：《浙江大學(xué)》2012年博士論文

【摘要】：移動(dòng)電話射頻電磁場(chǎng)的遺傳毒性效應(yīng)是其健康危險(xiǎn)度評(píng)估中的關(guān)鍵科學(xué)問(wèn)題。由于所采用的研究模型、輻照參數(shù)、分析方法等不同,目前有關(guān)射頻電磁場(chǎng)遺傳毒性的研究結(jié)果不僅很不一致,而且缺乏可比性,無(wú)法得出定論。有證據(jù)顯示,電磁場(chǎng)的生物學(xué)效應(yīng)與所研究的生物學(xué)系統(tǒng)的遺傳背景相關(guān)。我們認(rèn)為,在研究電磁場(chǎng)遺傳毒性效應(yīng)時(shí),首先應(yīng)該使用能指征電磁場(chǎng)這種弱作用因素對(duì)DNA影響的靈敏方法,系統(tǒng)地比較不同來(lái)源細(xì)胞對(duì)電磁場(chǎng)的響應(yīng)情況,即確定電磁場(chǎng)敏感細(xì)胞；然后,以敏感細(xì)胞為研究對(duì)象,深入探索電磁場(chǎng)的遺傳毒性和其它生物學(xué)效應(yīng)。 在本學(xué)位論文中,我們首先采用指示DNA雙鏈斷裂的早期、靈敏、特異的金指標(biāo)一一yH2AX焦點(diǎn)形成分析技術(shù),以六種不同系統(tǒng)來(lái)源的代表性細(xì)胞為研究對(duì)象,檢測(cè)移動(dòng)電話射頻電磁場(chǎng)對(duì)DNA雙鏈斷裂的影響,以明確射頻電磁場(chǎng)的遺傳毒性作用以及該作用是否具有細(xì)胞種類依賴性；如發(fā)現(xiàn)敏感細(xì)胞,則以此為研究對(duì)象,進(jìn)一步分析其后續(xù)生物學(xué)效應(yīng)。 結(jié)果顯示,比吸收率(SAR)為3W/kg的1800MHz射頻電磁場(chǎng)間斷(5minon/10min off)輻照細(xì)胞1小時(shí),沒(méi)有引起中國(guó)倉(cāng)鼠肺成纖維細(xì)胞(CHL)、SD大鼠星形膠質(zhì)細(xì)胞(Astrocytes)、人羊膜上皮細(xì)胞(FL)、人臍靜脈內(nèi)皮細(xì)胞(HUVEC)、人眼晶狀體上皮細(xì)胞(HLEC)和人皮膚成纖維細(xì)胞(HSF)六種細(xì)胞的DNA雙鏈斷裂；間斷輻照24小時(shí),僅觀察到CHL和HSF細(xì)胞yH2AX焦點(diǎn)數(shù)的顯著增加。 在以上發(fā)現(xiàn)射頻電磁場(chǎng)輻照致HSF細(xì)胞γH2AX焦點(diǎn)增加的基礎(chǔ)上,我們采用彗星實(shí)驗(yàn)、PI染色結(jié)合細(xì)胞流式檢測(cè)、細(xì)胞計(jì)數(shù)、CCK-8檢測(cè)、DCFH-DA探針?lè)跤Y(jié)合細(xì)胞流式檢測(cè)和免疫熒光成像等方法分析了其后續(xù)生物學(xué)效應(yīng)。結(jié)果顯示,射頻電磁場(chǎng)誘導(dǎo)的HSF細(xì)胞γH2AX焦點(diǎn)增加未能引起細(xì)胞內(nèi)DNA斷裂片段以及細(xì)胞周期、細(xì)胞增殖和細(xì)胞活力等的顯著改變；沒(méi)有同步改變?chǔ)肏2AX焦點(diǎn)復(fù)合物中的53BP1和Rad51焦點(diǎn)的形成；也沒(méi)有引起HSF細(xì)胞內(nèi)ROS水平的顯著變化。 根據(jù)以上研究結(jié)果,我們得出如下結(jié)論：1)在本實(shí)驗(yàn)條件下,移動(dòng)電話射頻電磁場(chǎng)以時(shí)間相關(guān)和細(xì)胞種類依賴的方式誘導(dǎo)細(xì)胞γH2AX焦點(diǎn)的增加；2)CHL和HSF細(xì)胞是響應(yīng)移動(dòng)電話射頻電磁場(chǎng)輻照的相對(duì)敏感細(xì)胞；3)移動(dòng)電話射頻電磁場(chǎng)誘導(dǎo)的γH2AX焦點(diǎn)增加并不導(dǎo)致顯著的生物學(xué)后果。 本學(xué)位論文的主要?jiǎng)?chuàng)新點(diǎn)：研究思路和方法方面,1)首次采用DNA損傷檢測(cè)的靈敏新方法,系統(tǒng)評(píng)價(jià)射頻電磁場(chǎng)的遺傳毒性效應(yīng)；2)首次以篩選出的射頻電磁場(chǎng)敏感細(xì)胞為研究對(duì)象,系統(tǒng)分析其后續(xù)生物學(xué)效應(yīng)。在科學(xué)研究方面,1)首次報(bào)道CHL和HSF細(xì)胞為射頻電磁場(chǎng)相對(duì)敏感細(xì)胞；2)首次報(bào)道了射頻電磁場(chǎng)誘導(dǎo)的γH2AX焦點(diǎn)增加未能引起細(xì)胞基因組不穩(wěn)定等后續(xù)生物學(xué)效應(yīng)的發(fā)生。
[Abstract]:Genetic toxicity of radio frequency electromagnetic field (RF EMF) in mobile phones is a key scientific problem in health risk assessment. Due to the different models, radiation parameters and analytical methods, the current research results on genetic toxicity of radiofrequency electromagnetic field are not only inconsistent, but also lack comparability, so it is impossible to draw a conclusion. There is evidence that the biological effects of electromagnetic fields are related to the genetic background of the biological systems studied. We believe that in studying the genotoxic effects of electromagnetic fields, we should first use a sensitive method that can indicate the effects of electromagnetic fields on DNA, and systematically compare the responses of cells from different sources to electromagnetic fields. That is, to determine the electromagnetic field sensitive cells, and then to explore the genotoxicity and other biological effects of the electromagnetic field by taking the sensitive cells as the research object. In this dissertation, we first used the yH2AX focus formation analysis technique, which indicates the early, sensitive and specific gold index of DNA double strand breaks, and studied the representative cells from six different systems. To detect the effects of radio frequency electromagnetic fields on DNA double strand breaks in mobile phones in order to determine the genotoxicity of radio frequency electromagnetic fields and whether they are cell type dependent and, if sensitive cells are found, to do so. Further analysis of its subsequent biological effects. The results showed that the specific absorptivity of 3W/kg was 5 min / 10 min off of the 1800MHz radiofrequency electromagnetic field (1800MHz) irradiation for 1 hour. The DNA double strand breaks of six kinds of cells, such as Astrocytes, human amniotic membrane epithelial cells, human umbilical vein endothelial cells, human ocular lens epithelial cells and human skin fibroblasts, have not been caused by the Chinese hamster lung fibroblasts (CHL) and SD rat astrocytes. After 24 hours of intermittent irradiation, only a significant increase in the number of yH2AX focal points in CHL and HSF cells was observed. On the basis of the above finding that the 緯 H2AX focus of HSF cells was increased by radiofrequency electromagnetic field irradiation, we used comet assay and Pi staining combined with flow cytometry. The subsequent biological effects were analyzed by cell counting and CCK-8 detection with DCFH-DA probe incubation combined with cell flow detection and immunofluorescence imaging. The results showed that the increase of 緯 H2AX focus in HSF cells induced by radiofrequency electromagnetic field did not cause significant changes in DNA fragmentation, cell cycle, cell proliferation and cell viability. The formation of 53BP1 and Rad51 focal points in 緯 H2AX focal complexes was not changed synchronously, nor did it cause significant changes in ROS levels in HSF cells. Based on the above results, we draw the following conclusion: 1) under the present experimental conditions, Radio frequency electromagnetic field of mobile phone induces increase of cell 緯 H2AX focal point in a time dependent and cell type dependent manner. HSF and HSF cells are relatively sensitive cells in response to radio frequency electromagnetic field irradiation of mobile phone.) Radio frequency electromagnetic field of mobile phone The induced increase in 緯 H2AX focus did not result in significant biological consequences. The main innovation of this dissertation is: research ideas and methods: for the first time, a new sensitive method of DNA damage detection is used to systematically evaluate the genotoxicity of radiofrequency electromagnetic field (RF-EMF) sensitive cells are used as the research object for the first time. The following biological effects were systematically analyzed. For the first time, we report that CHL and HSF cells are relatively sensitive to radiofrequency electromagnetic field (RF-EMF). It is the first time to report that the increase of 緯 H2AX focus induced by RF-EMF does not lead to the subsequent biological effects such as genomic instability.
【學(xué)位授予單位】：浙江大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2012
【分類號(hào)】：R363

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本文編號(hào)：1938089