【摘要】：背景 隨著的移動(dòng)通訊設(shè)備的廣泛使用,射頻電磁輻射可能帶來(lái)的不良效應(yīng)越來(lái)越受到大眾的關(guān)注。氧化應(yīng)激系體內(nèi)氧化與抗氧化失衡從而導(dǎo)致了機(jī)體生理生化改變和多種慢性疾病,因此射頻電磁輻射與氧化應(yīng)激的關(guān)系近年來(lái)已成為人們的研究方向。然而低強(qiáng)度射頻電磁輻射能否誘導(dǎo)氧化應(yīng)激目前還存在爭(zhēng)議,且射頻電磁輻射誘導(dǎo)氧化應(yīng)激的機(jī)制也不清楚。已知很多眼科疾病和氧化應(yīng)激有關(guān),射頻電磁輻射誘發(fā)晶狀體上皮細(xì)胞的生物學(xué)效應(yīng)也有大量報(bào)道。因此,研究低強(qiáng)度射頻電磁輻射是否能誘導(dǎo)晶狀體上皮細(xì)胞氧化應(yīng)激,并且對(duì)誘導(dǎo)氧化應(yīng)激的機(jī)理做進(jìn)一步研究,將有助于了解射頻電磁輻射所致晶狀體上皮細(xì)胞損傷的機(jī)制。 目的 探討1.8GHz射頻電磁輻射對(duì)體外培養(yǎng)的人晶狀體上皮細(xì)胞氧化應(yīng)激的影響以及可能的機(jī)制。 方法 體外培養(yǎng)人晶狀體上皮細(xì)胞(hLECs)分為輻照組與假輻照組。同時(shí)將其置于1.8GHz射頻電磁場(chǎng)內(nèi)進(jìn)行間斷(5分鐘開,10分鐘停)輻照,輻照組輻照強(qiáng)度比吸收率(specific absorption rate, SAR)為2,3,或4W/kg,假輻照為0W/kg。經(jīng)過(guò)0.5,1,或1.5小時(shí)輻照后,立刻使用熒光探針2’,7’-二氯熒光素二乙酸酯(2',7'-dichlorofluorescin diacetate, DCFH-DA)檢測(cè)細(xì)胞內(nèi)活性氧(reactive oxygen species, ROS)水平。經(jīng)過(guò)6,12,或24小時(shí)輻照后,使用丙二醛(malondialdehyde, MDA)檢測(cè)試劑盒測(cè)量細(xì)胞脂質(zhì)過(guò)氧化水平。經(jīng)過(guò)6,12,或24小時(shí)輻照后,使用CCK-8(Cell Counting Kit-8)試劑盒檢測(cè)細(xì)胞活力。 運(yùn)用實(shí)時(shí)定量PCR技術(shù)在轉(zhuǎn)錄水平研究1.8GHz射頻電磁輻射對(duì)抗氧化基因SOD1, SOD2, CAT和GPX1表達(dá)的影響。人晶狀體上皮細(xì)胞接受1小時(shí)輻照,提取總RNA經(jīng)逆轉(zhuǎn)錄后用實(shí)時(shí)熒光定量PCR試驗(yàn)檢測(cè)上述4個(gè)抗氧化基因的表達(dá)水平。 運(yùn)用Western blot技術(shù)在蛋白水平研究1.8GHz射頻電磁輻射對(duì)抗氧化酶SOD1, SOD2, CAT和GPx1的影響。人晶狀體上皮細(xì)胞接受1小時(shí)輻照,提取總蛋白后用使用Western blot檢測(cè)上述4個(gè)抗氧化酶蛋白的表達(dá)水平。 結(jié)果 經(jīng)過(guò)0.5,1,或1.5小時(shí)1.8GHz射頻電磁場(chǎng)輻照后,輻照組晶狀體上皮細(xì)胞內(nèi)的ROS水平明顯高于假輻照組(P0.05)。經(jīng)過(guò)6,12,或24小時(shí)輻照后,輻照組細(xì)胞培養(yǎng)基中的MDA濃度水平明顯高于假輻照組(P0.05)。經(jīng)過(guò)6,12,或24小時(shí)輻照后,輻照組細(xì)胞細(xì)胞活力明顯低于假輻照組(P0.05)。實(shí)時(shí)定量RT-PCR結(jié)果顯示經(jīng)過(guò)1小時(shí)輻照,輻照組細(xì)胞SOD1, SOD2, CAT和GPX1的mRNA表達(dá)顯著低于假輻照組(P0.05)。Western blot試驗(yàn)顯示經(jīng)過(guò)1小時(shí)輻照,輻照組細(xì)胞SOD1, SOD2,CAT和GPx1蛋白表達(dá)明顯低于假輻照組(P0.05)。 結(jié)論 低強(qiáng)度1.8GHz射頻電磁輻射能使得晶狀體上皮細(xì)胞產(chǎn)生氧化應(yīng)激。細(xì)胞內(nèi)ROS水平上升可能和射頻電磁輻射下抗氧化酶基因下調(diào)有關(guān)。
[Abstract]:Background with the wide use of mobile communication devices, the potential adverse effects of RF electromagnetic radiation have attracted more and more attention. Oxidative stress is the imbalance between oxidation and oxidation in vivo, which leads to physiological and biochemical changes and many chronic diseases. Therefore, the relationship between radiofrequency electromagnetic radiation and oxidative stress has become a research direction in recent years. However, whether low intensity radiofrequency electromagnetic radiation can induce oxidative stress is still controversial, and the mechanism of radiofrequency electromagnetic radiation induced oxidative stress is not clear. Many eye diseases are known to be related to oxidative stress, and the biological effects of radiofrequency electromagnetic radiation on lens epithelial cells have also been reported. Therefore, to study whether low intensity radiofrequency electromagnetic radiation can induce oxidative stress of lens epithelial cells, and to further study the mechanism of inducing oxidative stress, will be helpful to understand the mechanism of lens epithelial cell injury induced by radio frequency electromagnetic radiation. Objective to investigate the effect of 1.8GHz radiofrequency electromagnetic radiation on oxidative stress in cultured human lens epithelial cells and its possible mechanism. Methods cultured human lens epithelial cells (hLECs) were divided into irradiation group and false irradiation group. At the same time, it was placed in the 1.8GHz radio frequency electromagnetic field for intermittent irradiation (5 minutes open, 10 minutes stop). The radiation intensity of the irradiation group was 2W / kg, or 4W / kg, and the radiation intensity was 0 W / kg. The specific absorptivity of the irradiation group was 2W / kg, or 4W / kg. After 0. 5 or 1. 5 hours of irradiation, the level of reactive oxygen species (Ros) (reactive oxygen species, ROS) in the cells was detected by fluorescence probe 2H 7G-dichlorofluorescin diacetate, DCFH-DA. The lipid peroxidation level was measured by malondialdehyde (MDA) (malondialdehyde, MDA) assay kit after irradiation for 6 ~ 12, or 24 hours. Cell viability was detected by CCK-8 (Cell Counting Kit-8 kit after irradiation for 6 ~ 12, or 24 hours. The effect of 1.8GHz radiofrequency electromagnetic radiation on the expression of antioxidant genes SOD1, SOD2, CAT and GPX1 was studied at transcriptional level by real-time quantitative PCR. Human lens epithelial cells (LECs) were irradiated for 1 hour. The total RNA was extracted by reverse transcription and the expression levels of the above four antioxidant genes were detected by real-time fluorescence quantitative PCR assay. The effects of 1.8GHz RF electromagnetic radiation on SOD1, SOD2, CAT and GPx1 were studied at protein level by Western blot technique. Human lens epithelial cells were irradiated for 1 hour. The total protein was extracted and the expression of the above four antioxidant enzyme proteins was detected by Western blot. Results the ROS level of lens epithelial cells in the irradiated group was significantly higher than that in the sham irradiation group (P0.05). The MDA concentration in the cell culture medium of the irradiated group was significantly higher than that of the sham irradiation group (P0.05). The cell viability in the irradiation group was significantly lower than that in the sham irradiation group (P0.05). The results of real-time quantitative RT-PCR showed that after 1 hour irradiation, the mRNA expression of SOD1, SOD2, CAT and GPX1 in irradiated cells was significantly lower than that in false irradiation group (P0.05). Western blot test showed that after 1 hour irradiation, the SOD1, SOD2, expression of irradiated cells was significantly lower than that of false irradiation group). The expression of CAT and GPx1 protein was significantly lower than that of false irradiation group (P0.05). Conclusion low intensity 1.8GHz radiofrequency electromagnetic radiation can induce oxidative stress in lens epithelial cells. The increase of intracellular ROS level may be related to the down-regulation of antioxidant enzyme gene under radiofrequency electromagnetic radiation.
【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】：2013
【分類號(hào)】：R779.1

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