Interaction of oxygen-sensitive luminescent probes Ru(phen)32+ and Ru(bipy)32+ with animal and plant cells in vitro -: Mechanism of phototoxicity and conditions for non-invasive oxygen measurements
Interaction of oxygen-sensitive luminescent probes Ru(phen)32+ and Ru(bipy)32+ with animal and plant cells in vitro -: Mechanism of phototoxicity and conditions for non-invasive oxygen measurements
复制标题
DOI：
10.1016/s1011-1344(01)00257-3
复制
发表时间：
2001-12-31
影响因子：
5.4
通讯作者：
Dobrucki, JW
中科院分区：
文献类型：
--
作者：
Dobrucki, JW
了解氧在动物细胞的生理学、病理生理学以及放射和化学敏感性中的作用，需要在体外或体内对细胞中0- 2X 10（-4）M范围内的氧浓度进行准确和非侵入性的测量。高分辨率3D成像技术在研究体内组织氧合和体外模型组织（多细胞球体）中可能特别有用。本工作的目标是发展显微技术和（i）确定两种氧敏发光染料Ru（bipy）（3）（2-）（三（2，2 '-联吡啶）氯化钌（II）六水合物）和Ru（phen）（3）（2-）（尿（1，10-菲咯啉）氯化钌（II）水合物）可用于探测体外活细胞内的氧浓度，当没有明显的光毒性作用时。以及（ii）一旦细胞损伤发生，研究光毒性的机制。本报告表明，Ru（bipy）（3）（2-）和Ru（phen）（3）（2-）不会穿过完整的生物膜，在0.2 mM的浓度下不会对质膜造成可测量的光损伤，并且当加载到内体中时，产生强烈的发光信号。然而，在1 mM的细胞外浓度下，在457 nm光的存在下，可检测量的两种复合物在质膜上积累，并通过可能涉及单线态氧产生的机制导致膜完整性的丧失。(C)2001 Elsevier Science B. V.保留所有权利。
Understanding the role of oxygen in the physiology, pathophysiology and radio- and chemosensitivity of animal cells requires accurate and non-invasive measurements of oxygen concentrations in the range of 0-2X10(-4) M, in cells in vitro or in vivo. High resolution 3D imaging techniques could be particularly useful in investigating tissue oxygenation in vivo and in model tissues (multicellular spheroids) in vitro. The goals of this work were to develop microscopy techniques and (i) to define conditions under which two oxygen-sensitive luminescent dyes, Ru(bipy)(3)(2-) (tris(2,2'-bipyridyl)ruthenium(II) chloride hexahydrate) and Ru(phen)(3)(2-) (uris(1,10-phenanthroline)ruthenium(II) chloride hydrate) can be used to probe oxygen concentrations within viable cells in vitro, when no phototoxic effects are evident. and (ii) to investigate the mechanism of phototoxicity once cell damage occurs. This report demonstrates that Ru(bipy)(3)(2-) and Ru(phen)(3)(2-) do not pass through intact biological membranes, do not cause measurable photodamage to plasma membranes at a concentration of 0.2 mM and, when loaded into endosomes, yield a strong luminescent signal. However, at an extracellular concentration of 1 mM, in the presence of 457-nm light, detectable amounts of both complexes accumulate at the plasma membrane and cause a loss of membrane integrity via a mechanism which may involve the generation of singlet oxygen. (C) 2001 Elsevier Science B.V. All rights reserved.