Santini M T, Ferrante A, Romano R, Rainaldi G, Motta A, Donelli G, Vecchia P, Indovina P L
Dipartimento di Ematologia, Oncologia e Medicina Molecolare, Istituto Superiore di Sanità, Rome, Italy.
Int J Radiat Biol. 2005 Feb;81(2):97-113. doi: 10.1080/09553000500102985.
To study cell damage and possible apoptosis in K562 human erythroleukemic cells exposed for 2 h to an extremely low frequency (ELF) 50 Hz sinusoidal magnetic field with a magnetic induction of either 1 or 5 mT using high resolution proton nuclear magnetic resonance (1H-NMR) spectroscopy.
One-dimensional 1H-NMR spectra were obtained on whole K562 cells and perchloric acid extracts of these cells. In addition, two-dimensional 1H-NMR spectra were also acquired. Cell damage was examined by lactate dehydrogenase release and changes in cell growth were monitored by growth curve analyses, bromodeoxyuridine incorporation and Ki67 antigen localization. Cell death (necrosis and apoptosis) were also studied by using the chromatin dye Hoechst 33258.
The variations in numerous metabolites observed with 1H-NMR reveal apoptosis-like behavior in response of K562 cells to ELF fields.
1H-NMR can be extremely useful in studying the effects of ELF fields on cells. In particular, the variations in metabolites which suggest apoptosis-like behavior occur when the cells are not identifiable as apoptotic by more traditional techniques.
利用高分辨率质子核磁共振(1H-NMR)光谱法,研究暴露于磁感应强度为1或5 mT的50 Hz极低频(ELF)正弦磁场2小时的K562人红白血病细胞中的细胞损伤及可能的凋亡情况。
获取了完整K562细胞及其高氯酸提取物的一维1H-NMR光谱。此外,还采集了二维1H-NMR光谱。通过乳酸脱氢酶释放检测细胞损伤,并通过生长曲线分析、溴脱氧尿苷掺入及Ki67抗原定位监测细胞生长变化。还使用染色质染料Hoechst 33258研究细胞死亡(坏死和凋亡)情况。
1H-NMR观察到的多种代谢物变化揭示了K562细胞对ELF场反应中的凋亡样行为。
1H-NMR在研究ELF场对细胞的影响方面极为有用。特别是,当细胞通过更传统技术无法鉴定为凋亡时,代谢物的变化表明存在凋亡样行为。