臺灣博碩士論文加值系統

摘要
生物體中幾乎所有生理表現均由微小的電磁訊號傳遞來完成，而生物體中分子的鍵結其能量均遠大於微波及極低頻電磁場的能量。該電磁場之能量雖不足以打斷其鍵結，但對這些具有電子的分子會有極化、飄移等作用，因此可能對細胞造成影響。
今我們先以鈷60或過氧化氫在溶液中所產生的自由基來攻擊V79-4細胞的DNA，使細胞關於修復DNA的基因能夠表現而進行修復DNA的工作。在修復的同時，我們給予不同的電磁波照射；之後利用MTT比色分析法來觀察細胞的存活率。MTT比色分析法的原理是利用MTT藥劑會與粒線體上電子傳遞鏈中的電子產生作用，使其顏色由黃轉紫，測其OD值以換算成細胞存活率。實驗結果顯示，不論是以鈷60或是過氧化氫所傷害的細胞，在修復DNA的同時若以極低頻電磁場加以照射，並不會影響其存活率。但對DNA已受到傷害的細胞加以照射微波波段電磁波時，經過統計的檢定後發現存活率有明顯的下降趨勢。為確認細胞是在受到DNA傷害後其存活率才會受到電磁波的影響，將細胞直接以不同頻率的電磁波照射而加以觀察，並未發現其存活率有明顯變化。
另外我們也使用單細胞電泳法來觀察細胞中DNA受損的情形。因本次實驗無法觀察小劑量鈷60照射或是過氧化氫的傷害，所以將來改進之道為找到更好的實驗條件，如電泳時間、電泳液的成分等以利用電泳法觀察細胞中DNA受損情形。

Abstract
All functions in biological system are almost accomplished by transduction of electromagnetic signal; the binding energy among these biological molecules is larger than that of microwaves and extremely low frequency(ELF) electromagnetic(EM) fields. The energy of EM waves is incapable of breaking the bindings, but affect the electrons within the molecules for their polarization and drift, etc. So the cell properties may be influenced by the EM waves.
In this dissertation, we utilized the free radicals that were generated by from Co-60 or hydrogen peroxide to attack the DNA of the V79-4 cell. The DNA repair-gene was triggered to repair the free radical damaged DNA. Meanwhile, The EM waves irradiated the target cell. The 2-[4,5— Dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide (MTT) assay presented the surviving fraction of the cell. The MTT powder changed the color from yellow to purple by an interaction between the reagent and the electrons in the electron-transport chain on the mitochondria. The optical density (OD) value retrieved from MTT assay deduced the surviving fraction of the cell. The results indicated that the ELF EM wave had no influences on the surviving fraction of the cells that were damaged by Co-60 or hydrogen peroxide. However, when the microwaves irradiated on the DNA-damaged cells, the statistical test clarified that the surviving fraction of the cell decreased significantly. In order to confirm the DNA-damaged cells were sensitive to EM waves, the microwaves and ELF EM waves were irradiating the cells that the DNA was not damaged. After obtaining the surviving fraction of the undamaged cell the experiments indicated that surviving fraction had no significant variation.
In addition, we measure the DNA damages by single cell gel electrophoresis method. Nonetheless, the slight dose irradiator of Co-60 or hydrogen peroxide performed no effect in the strategy. The experimental conditions, such as the electrophoresis time and running buffers, could be further optimized in the future research.

目錄
中文摘要Ⅰ
英文摘要Ⅱ
致謝Ⅲ
目錄 目錄Ⅰ
圖目錄與表目錄目錄Ⅳ
第一章、緒論1
1.1 文獻回顧1
1.2 研究方向與目的6
第二章、原理8
2.1 電磁波的生物效應8
2.1.1電磁波的微觀效應8
2.1.2 電磁波的中觀效應9
2.1.3 電磁波的巨觀效應10
2.2 微波波段電磁波生物效應11
2.2.1 熱效應11
2.2.2 非熱效應12
2.3 極低頻電磁場生物效應13
2.4 電磁波對生物作用機制的假設13
2.5 生物實驗原理14
2.5.1 過氧化氫對細胞的傷害14
2.5.2 鈷60對細胞的傷害15
2.5.3 比色分析法(MTT Assay)15
2.5.4 單細胞電泳分析法(Single Cell Gel Electrophoresis)16
2.5.5 分次劑量效應17
第三章、實驗系統與設計18
3.1 實驗設備18
3.1.1 微波波段電磁波18
3.1.1.1 微波共振腔19
3.1.1.2 微波電磁場量測儀器20
3.1.2 極低頻波段電磁場21
3.1.2.1 Helmholtz線圈照射場21
3.1.2.2極低頻電磁場量測儀器27
3.2 游離輻射照射場28
3.3 生物材料28
3.3.1 細胞株(Cell line)28
3.3.2 化學藥品與試劑29
3.4 實驗方法30
3.4.1 比色分析法30
3.4.1.1 細胞存活率實驗31
3.4.1.2電磁場對細胞修復DNA影響之實驗34
3.4.2 單細胞電泳分析法36
第四章、實驗結果與討論38
4.1 電磁場量測38
4.1.1 微波波段電磁波38
4.1.2 極低頻電磁場電磁波41
4.2 微波波段電磁波共振腔GTEM 250之溫度控制43
4.3 SAR量測46
4.4比色分析法48
4.4.1 微波波段及極低頻電磁場對細胞存活率的影響48
4.4.2 過氧化氫對細胞存活率的影響50
4.4.2.1 不同濃度之過氧化氫影響下的細胞存活率50
4.4.2.2 分次劑量效應51
4.4.2.3 微波電磁波與極低頻電磁場對經過過氧化氫
傷害的細胞其存活率的影響51
4.4.3 鈷60放射線對細胞存活率的影響53
4.4.3.1 不同劑量之鈷60放射線影響下的細胞存活率54
4.4.3.2 分次劑量效應54
4.4.3.3 微波電磁波與極低頻電磁場對經過鈷60放射
線傷害的細胞其存活率的影響55
4.5 單細胞電泳分析法57
第五章、結論58
參考文獻61
附錄一、單細胞電泳法之亮度質心分析65
附錄二、細胞懸服役之比熱量測69
附錄三、兩個母群體t-TEST檢定方法71
圖表目錄
圖2.1 時變電磁場對細胞的效應8
圖2.2 肌肉組織介電係數和導電常數隨電磁波頻率f的變化圖9
圖2.3 DNA受到游離輻射攻擊的機制15
圖2.4 以Scrolapp軟體所定義之彗星頭部與彗星長度
之示意圖16
圖2.5 單次與分次劑量對細胞存活率之影響17
圖3.1 微波照射的實驗裝置圖19
圖3.2 GTEM微波共振腔內部構造圖20
圖3.3 EMR-30電場量測棒21
圖3.4單匝圓形線圈22
圖3.5 Helmholtz 線圈22
圖3.6 Helmholtz 線圈-球座標25
圖3.7 Helmholtz 線圈裝置圖26
圖3.8高斯磁場分佈模擬結果27
圖3.9 EFA-3 低頻電磁場量測儀器27
圖3.10鈷60放射源之劑量率與距離關係圖28
圖3.11 96-well microplate示意圖33
圖3.12鈷60照射示意圖33
圖4.1自製鋁門洞口漏電場功率密度對時間的變化39
圖4.2共振腔內一點電場功率密度對時間的變化39
圖4.3 微波共振腔測試區強度分佈圖40
圖4.4 Helmoltz線圈磁場強度42
圖4.5 Helmoltz線圈背景值42
圖4.6 高斯磁場分佈圖43
圖4.7 微波共振腔溫度控制示意圖44
圖4.8 微波共振腔溫度控制反應圖45
圖4.9 經溫度控制後，細胞懸浮液的增溫情形46
圖4.10 微波電磁波對細胞懸浮液增溫關係圖47
圖4.11 受到10G低頻電磁場照射後的細胞存活率48
圖4.12 受到5G低頻電磁場照射後的細胞存活率49
圖4.13 受到2.45GHz電磁波照射後的細胞存活率49
圖4.14 V79-4細胞對不同濃度的過氧化氫存活曲線圖50
圖4.15 V79-4細胞單次與分次接受過氧化氫傷害之存活
率曲線51
圖4.16 V79-4細胞經由過氧化氫處理後，再照射10Gauss
極低頻電磁場之存活率曲線52
圖4.17 V79-4細胞經由過氧化氫處理後，再照射5Gauss
極低頻電磁場之存活率曲線52
圖4.18 V79-4細胞經由過氧化氫處理後，再照射微低之
存活率曲線53
圖4.19 V79-4細胞對鈷60照射劑量的存活曲線54
圖4.20 V79-4細胞單次與分次接受鈷60照射之存活率曲線55
圖4.21 V79-4細胞經由鈷60照射後，再照射10Gauss
極低頻電磁場之存活率曲線55
圖4.22 V79-4細胞經由鈷60照射後，再照射5Gauss
極低頻電磁場之存活率曲線56
圖4.23 V79-4細胞經由鈷60照射後，再照射微低之
存活率曲線56
圖A.1 單細胞電泳圖及定義區域66
圖A.2 不同劑量的單細胞電泳圖66
圖A.3 尾部長度與彗星尾部質心比較圖67
圖A.4 使用過氧化氫和鈷60傷害細胞DNA的劑量
反應關係圖68
圖C.1 獨立樣本分為兩群，各群規模為371
圖C.2 成對樣本設計，包含三組實驗單位72
表2.1 肌肉組織受到不同頻率外加電場造成介電常數及
導電係數改變一覽表10
表3.1 Microplate內不同格所對應的位置和劑量34
表4.1利用熱平衡的方法量測細胞懸浮液的比熱47
表4.2 不同電磁波對細胞在存活率上的影響49
表4.3過氧化氫處理後以不同電磁波影響細胞修復DNA
的存活率，以t-TEST檢定所得的p值53
表4.4 經過鈷60照射後以不同電磁波影響細胞修復DNA
的存活率，以t-TEST檢定所得的p值57
表B.1 利用熱平衡的方法量測純水的比熱70

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