Joubert Vanessa, Bourthoumieu Sylvie, Leveque Philippe, Yardin Catherine
Department of Cell Biology, Histology and Cytogenetics, EA3842 Limoges University Hospital, Faculty of Medicine, 87025 Limoges Cedex, France.
Radiat Res. 2008 Jan;169(1):38-45. doi: 10.1667/RR1077.1.
Joubert, V., Bourthoumieu, S., Leveque, P. and Yardin, C. Apoptosis is Induced by Radiofrequency Fields through the Caspase-Independent Mitochondrial Pathway in Cortical Neurons. Radiat. Res. 169, 38-45 (2008). In the present study, we investigated whether continuous-wave (CW) radiofrequency (RF) fields induce neuron apoptosis in vitro. Rat primary neuronal cultures were exposed to a CW 900 MHz RF field with a specific absorption rate (SAR) of 2 W/kg for 24 h. During exposure, an increase of 2 degrees C was measured in the medium; control experiments with neurons exposed to 39 degrees C were then performed. Apoptosis was assessed by condensation of nuclei with 4',6-diamino-2-phenylindole (DAPI) staining observed with an epifluorescence microscope and fragmentation of DNA with TdT-mediated dUTP nick-end labeling (TUNEL) analyzed by flow cytometry. A statistically significant difference in the rate of apoptosis was found in the RF-field-exposed neurons compared to the sham-, 37 degrees C- and 39 degrees C-exposed neurons either 0 or 24 h after exposure using both methods. To assess whether the observed apoptosis was caspase-dependent or -independent, assays measuring caspase 3 activity and apoptosis-inducing factor (AIF) labeling were performed. No increase in the caspase 3 activity was found, whereas the percentage of AIF-positive nuclei in RF-field-exposed neurons was increased by three- to sevenfold compared to other conditions. Our results show that, under the experimental conditions used, exposure of primary rat neurons to CW RF fields may induce a caspase-independent pathway to apoptosis that involves AIF.
朱伯特,V.,布尔图米厄,S.,勒韦克,P.和雅丹,C. 射频场通过皮质神经元中不依赖半胱天冬酶的线粒体途径诱导细胞凋亡。《辐射研究》169卷,第38 - 45页(2008年)。在本研究中,我们调查了连续波(CW)射频(RF)场是否在体外诱导神经元凋亡。将大鼠原代神经元培养物暴露于比吸收率(SAR)为2 W/kg的900 MHz连续波射频场中24小时。暴露期间,培养基温度升高了2摄氏度;随后对暴露于39摄氏度的神经元进行对照实验。通过用落射荧光显微镜观察4',6 - 二脒基 - 2 - 苯基吲哚(DAPI)染色检测细胞核凝聚,并通过流式细胞术分析末端脱氧核苷酸转移酶介导的dUTP缺口末端标记(TUNEL)检测DNA片段化来评估细胞凋亡。使用这两种方法,在暴露0小时或24小时后,与假暴露、37摄氏度暴露和39摄氏度暴露的神经元相比,在暴露于射频场的神经元中发现细胞凋亡率存在统计学显著差异。为了评估观察到的细胞凋亡是依赖半胱天冬酶还是不依赖半胱天冬酶,进行了测量半胱天冬酶3活性和凋亡诱导因子(AIF)标记的检测。未发现半胱天冬酶3活性增加,而与其他条件相比,暴露于射频场的神经元中AIF阳性细胞核的百分比增加了三到七倍。我们的结果表明,在所使用的实验条件下,将原代大鼠神经元暴露于连续波射频场可能诱导一条不依赖半胱天冬酶的细胞凋亡途径,该途径涉及AIF。