Metal-sulfate induced generation of ROS in human brain cells: detection using an isomeric mixture of 5- and 6-carboxy-2',7'-dichlorofluorescein diacetate (carboxy-DCFDA) as a cell permeant tracer.

Metal-sulfate induced generation of ROS in human brain cells: detection using an isomeric mixture of 5- and 6-carboxy-2',7'-dichlorofluorescein diacetate (carboxy-DCFDA) as a cell permeant tracer.
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DOI:
10.3390/ijms13089615
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发表时间:
2012
影响因子:
5.6
通讯作者:
Lukiw WJ
Lukiw WJ
中科院分区:
生物学2区
文献类型:
--
作者:
Pogue AI;Jones BM;Bhattacharjee S;Percy ME;Zhao Y;Lukiw WJ

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在神经组织的病理生理应激过程中产生的活性氧(ROS)的进化与几种进行性人类神经系统疾病的病因有关,包括阿尔茨海默病(AD)和肌萎缩性侧索硬化症(ALS)。在这篇简短的交流中,我们使用了5-(和6)-羧基-2 ',7 ' -二氯荧光素双乙酸酯(羧基- dcfda; C25H14Cl2O9; MW 529.3)的混合异构体,一种新型荧光指示剂,来评估在原代共培养的人类神经元-胶质(HNG)细胞中ROS的产生。我们利用12种与环境、工业和农业相关的二价和三价金属硫酸盐引入了病理应激,包括Al、Cd、Cu、Fe、Hg、Ga、Mg、Mn、Ni、Pb、Sn和Zn。在这个实验测试系统中,在所分析的所有金属硫酸盐中,硫酸铝显示出迄今为止最大的诱导细胞内ROS的能力。这些研究表明,使用羧基- h2dcfda二乙酸酯的异构体混合物作为一种新型的、高灵敏度的、持久的、细胞渗透的、基于荧光素的示踪剂,可以定量完整的、代谢的人脑细胞中的ROS生成,并分析不同金属硫酸盐对ROS生成和ROS介导的神经功能障碍的潜在表观遗传贡献。
Evolution of reactive oxygen species (ROS), generated during the patho-physiological stress of nervous tissue, has been implicated in the etiology of several progressive human neurological disorders including Alzheimer’s disease (AD) and amylotrophic lateral sclerosis (ALS). In this brief communication we used mixed isomers of 5-(and-6)-carboxy-2′,7′-dichlorofluorescein diacetate (carboxy-DCFDA; C25H14Cl2O9; MW 529.3), a novel fluorescent indicator, to assess ROS generation within human neuronal-glial (HNG) cells in primary co-culture. We introduced pathological stress using the sulfates of 12 environmentally-, industrially- and agriculturally-relevant divalent and trivalent metals including Al, Cd, Cu, Fe, Hg, Ga, Mg, Mn, Ni, Pb, Sn and Zn. In this experimental test system, of all the metal sulfates analyzed, aluminum sulfate showed by far the greatest ability to induce intracellular ROS. These studies indicate the utility of using isomeric mixtures of carboxy-H2DCFDA diacetates as novel and highly sensitive, long-lasting, cell-permeant, fluorescein-based tracers for quantifying ROS generation in intact, metabolizing human brain cells, and in analyzing the potential epigenetic contribution of different metal sulfates to ROS-generation and ROS-mediated neurological dysfunction.
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