【摘要】：目的工頻磁場是頻率為50Hz的極低頻磁場,隨著人們接觸頻率及強(qiáng)度的提高,工頻磁場的生物學(xué)效應(yīng)引起了研究者的極大關(guān)注,有關(guān)工頻磁場對細(xì)胞DNA損傷修復(fù)的影響成為評價(jià)其健康風(fēng)險(xiǎn)的關(guān)鍵事件之一,但迄今為止有關(guān)其遺傳損傷效應(yīng)的研究報(bào)道尚無確切定論。本研究采用DNA雙鏈斷裂的早期敏感指標(biāo)—γH2AX,初步評價(jià)了工頻磁場不同輻照強(qiáng)度和時(shí)間對三種不同系統(tǒng)或器官來源細(xì)胞的DNA損傷效應(yīng),以期為工頻磁場敏感細(xì)胞的篩選及其進(jìn)一步的生物學(xué)效應(yīng)研究奠定基礎(chǔ)。 方法將人羊膜上皮細(xì)胞(FL)、人臍靜脈內(nèi)皮細(xì)胞(HUVEC)以及人皮膚成纖維細(xì)胞(HSF)分別暴露于50Hz,0.4mT、1mT和2mT工頻磁場15min、1h和24h后,采用γH2AX免疫熒光法和免疫印跡法分別檢測了細(xì)胞內(nèi)DNA雙鏈斷裂和γH2AX表達(dá)情況。每個(gè)強(qiáng)度和時(shí)間點(diǎn)分別設(shè)置一一對應(yīng)假暴露組,同時(shí)采用0.5μM或1μM4NQO處理1h作為陽性對照組。其中,γH2AX免疫熒光實(shí)驗(yàn)采用平均焦點(diǎn)數(shù)、焦點(diǎn)陽性細(xì)胞率以及焦點(diǎn)分段分布率作為DNA損傷程度的評價(jià)指標(biāo)。實(shí)驗(yàn)采用雙盲,每組實(shí)驗(yàn)重復(fù)3-5次。 結(jié)果γH2AX免疫熒光法和免疫印跡法結(jié)果顯示,工頻磁場各暴露組與一一對應(yīng)假暴露組相比,平均焦點(diǎn)數(shù)、焦點(diǎn)陽性細(xì)胞率、yH2AX蛋白表達(dá)量均沒有顯著性差異(P0.05),焦點(diǎn)分段分布率在個(gè)別強(qiáng)度或時(shí)間點(diǎn)上有差異(P0.05)。 結(jié)論在本實(shí)驗(yàn)條件下,工頻磁場未能導(dǎo)致FL、HUVEC以及HSF細(xì)胞DNA損傷。
[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, 緯 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 緯 H2AX in the cells were detected by 緯 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 緯 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 緯 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.
【學(xué)位授予單位】：浙江大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R142

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