【摘要】：背景 隨著的移動通訊設備的廣泛使用,射頻電磁輻射可能帶來的不良效應越來越受到大眾的關(guān)注。氧化應激系體內(nèi)氧化與抗氧化失衡從而導致了機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激的關(guān)系近年來已成為人們的研究方向。然而低強度射頻電磁輻射能否誘導氧化應激目前還存在爭議,且射頻電磁輻射誘導氧化應激的機制也不清楚。已知很多眼科疾病和氧化應激有關(guān),射頻電磁輻射誘發(fā)晶狀體上皮細胞的生物學效應也有大量報道。因此,研究低強度射頻電磁輻射是否能誘導晶狀體上皮細胞氧化應激,并且對誘導氧化應激的機理做進一步研究,將有助于了解射頻電磁輻射所致晶狀體上皮細胞損傷的機制。 目的 探討1.8GHz射頻電磁輻射對體外培養(yǎng)的人晶狀體上皮細胞氧化應激的影響以及可能的機制。 方法 體外培養(yǎng)人晶狀體上皮細胞(hLECs)分為輻照組與假輻照組。同時將其置于1.8GHz射頻電磁場內(nèi)進行間斷(5分鐘開,10分鐘停)輻照,輻照組輻照強度比吸收率(specific absorption rate, SAR)為2,3,或4W/kg,假輻照為0W/kg。經(jīng)過0.5,1,或1.5小時輻照后,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)檢測細胞內(nèi)活性氧(reactive oxygen species, ROS)水平。經(jīng)過6,12,或24小時輻照后,使用丙二醛(malondialdehyde, MDA)檢測試劑盒測量細胞脂質(zhì)過氧化水平。經(jīng)過6,12,或24小時輻照后,使用CCK-8(Cell Counting Kit-8)試劑盒檢測細胞活力。 運用實時定量PCR技術(shù)在轉(zhuǎn)錄水平研究1.8GHz射頻電磁輻射對抗氧化基因SOD1, SOD2, CAT和GPX1表達的影響。人晶狀體上皮細胞接受1小時輻照,提取總RNA經(jīng)逆轉(zhuǎn)錄后用實時熒光定量PCR試驗檢測上述4個抗氧化基因的表達水平。 運用Western blot技術(shù)在蛋白水平研究1.8GHz射頻電磁輻射對抗氧化酶SOD1, SOD2, CAT和GPx1的影響。人晶狀體上皮細胞接受1小時輻照,提取總蛋白后用使用Western blot檢測上述4個抗氧化酶蛋白的表達水平。 結(jié)果 經(jīng)過0.5,1,或1.5小時1.8GHz射頻電磁場輻照后,輻照組晶狀體上皮細胞內(nèi)的ROS水平明顯高于假輻照組(P0.05)。經(jīng)過6,12,或24小時輻照后,輻照組細胞培養(yǎng)基中的MDA濃度水平明顯高于假輻照組(P0.05)。經(jīng)過6,12,或24小時輻照后,輻照組細胞細胞活力明顯低于假輻照組(P0.05)。實時定量RT-PCR結(jié)果顯示經(jīng)過1小時輻照,輻照組細胞SOD1, SOD2, CAT和GPX1的mRNA表達顯著低于假輻照組(P0.05)。Western blot試驗顯示經(jīng)過1小時輻照,輻照組細胞SOD1, SOD2,CAT和GPx1蛋白表達明顯低于假輻照組(P0.05)。 結(jié)論 低強度1.8GHz射頻電磁輻射能使得晶狀體上皮細胞產(chǎn)生氧化應激。細胞內(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.
【學位授予單位】：浙江大學
【學位級別】：博士
【學位授予年份】：2013
【分類號】：R779.1

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