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2-NBDG,葡萄糖的荧光类似物,作为体外检测细胞电穿孔的标记物。

2-NBDG, a fluorescent analogue of glucose, as a marker for detecting cell electropermeabilization in vitro.

机构信息

Department of Medical Physics, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.

出版信息

J Membr Biol. 2012 Oct;245(10):633-42. doi: 10.1007/s00232-012-9479-6. Epub 2012 Aug 11.

Abstract

This study investigated whether molecules spontaneously transported inside cells, like glucose derivatives, can also be used as electropermeabilization markers. Uptake of a fluorescent deoxyglucose derivative (2-NBDG) by normal and electropermeabilized cells in culture was analyzed. 2-NBDG was added to DC-3F cell suspensions and cells, exposed or not to eight square-wave electric pulses of 100-μs duration and of appropriate field amplitude at a repetition frequency of 1 Hz or 5 kHz, were incubated at 37 °C. 2-NBDG uptake was temperature-, concentration- and time-dependent in cells submitted or not to the electric pulses. In spite of significant uptake of 2-NBDG mediated by GLUT transporters into nonpermeabilized cells, the electric pulses significantly increased about ten to hundred times the 2-NBDG uptake into the cells. The increase in the field amplitude from 900 to 1,500 V/cm resulted in a progressive increase of 2-NDBG. Our results show that under the conditions of in vivo exposure duration to FDG and the physiological concentration of D-glucose, electric pulses increased 2-NBDG uptake into electropermeabilized cells. Under our experimental conditions, the percentage of permeabilized cells within the population of cells exposed to electric pulses remained at the same level regardless of the pulse frequency used, 1 Hz or 5 kHz. The findings showed that glucose derivatives can also be used to detect electropermeabilized cells exposed to electric pulses.

摘要

本研究旨在探讨像葡萄糖衍生物等自发内运的分子是否也可作为电穿孔标记物。分析了培养中正常和电穿孔细胞对荧光脱氧葡萄糖衍生物(2-NBDG)的摄取。将 2-NBDG 添加到 DC-3F 细胞悬浮液中,将细胞在 37°C 下孵育,同时使细胞暴露或不暴露于 8 个 100-μs 时长和适当场强的方波电脉冲,重复频率为 1 Hz 或 5 kHz。在未施加电脉冲的细胞中,2-NBDG 的摄取量随温度、浓度和时间的变化而变化。尽管 GLUT 转运体介导的 2-NBDG 大量摄取到未穿孔的细胞中,但电脉冲仍使 2-NBDG 摄取量增加了约 10 至 100 倍。将场强从 900 增加到 1500 V/cm,导致 2-NBDG 逐渐增加。我们的结果表明,在体内实际暴露于 FDG 的时间和 D-葡萄糖的生理浓度条件下,电脉冲可增加电穿孔细胞对 2-NBDG 的摄取。在我们的实验条件下,无论使用的脉冲频率为 1 Hz 还是 5 kHz,暴露于电脉冲的细胞群体中透化细胞的百分比保持不变。研究结果表明,葡萄糖衍生物也可用于检测暴露于电脉冲的电穿孔细胞。

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