Abstract

The structure of nitric oxide (NO) donors, paramagnetic dinitrosyl iron complexes with SH ligands (DNICs), contains two nitrosyl groups, NO and NO⁺, which are released from a DNIC upon degradation in the cell. The selective ability of diethyl dithiocarbamate (DETC) to bind NO was used in this work to study the functions of nitrosonium cation NO⁺ in the regulation of biofilm formation by nitric oxide donors. The combined treatment of Pseudomonas aeruginosa with DETC and NO donors, sodium nitrite NaNO₂ or binuclear DNIC with glutathione (B-DNIChglu), multiply reduced the biofilm formation relative to the control and single treatments with each compound. The biofilm formation depended on the ratio of components. At a tenfold excess of DETC against DNICglu, the formation rate decreased by three times, and under a fivefold excess, by 1.8 times. The stable [Fe²⁺–DETC] complex formed during the combined treatment functioned as an NO trap, leading to the block in the synthesis of the signaling regulator DNICglu from NO and iron and to the accumulation of nitrosonium cation NO⁺ in the cell. The maximum decrease in the biofilm formation rate was established in the option with successive treatment of cells with DETC with a lag period of 40 min after the introduction of the NaNO₂ donor and the formation in DNIC with thiosulfate ligands, universal NO signaling molecules of all biosystems, in the cells. The results obtained expand knowledge about the functions of nitrosonium cation and contribute to the disclosure of mechanisms of the toxic activity of DNIC donors. They correlate with our earlier results on NO⁺ inhibition of E. coli colony formation in experiments with DNICglu, as well as with the data of other researchers obtained on yeast and in MCF7 cancer cell culture.

中文翻译：

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Nitrosonium 阳离子 NO+ 抑制一氧化氮在调节铜绿假单胞菌生物膜形成中的作用

摘要

一氧化氮 (NO) 供体的结构，顺磁性二亚硝基铁与 SH 配体 (DNIC) 的配合物，包含两个亚硝基基团 NO 和 NO + ，它们在^细胞中降解时从 DNIC 中释放出来。本研究利用二乙基二硫代氨基甲酸酯 (DETC) 结合 NO 的选择性来研究亚硝阳离子 NO ⁺在一氧化氮供体调节生物膜形成中的功能。DETC和NO供体亚硝酸钠NaNO ₂联合治疗铜绿假单胞菌或双核 DNIC 与谷胱甘肽 (B-DNIChglu)，相对于对照和每种化合物的单一处理，成倍减少生物膜形成。生物膜的形成取决于组分的比例。在相对于 DNICglu 十倍过量的 DETC 下，形成率降低了三倍，在五倍过量的情况下，降低了 1.8 倍。在联合处理过程中形成的稳定 [Fe ²⁺ –DETC] 复合物起到 NO 陷阱的作用，导致信号调节剂 DNICglu 从 NO 和铁的合成受阻，并导致细胞中亚硝阳离子 NO ⁺的积累。生物膜形成率的最大降低是在引入 NaNO 后用 DETC 连续处理细胞的选项中确定的，滞后期为 40 分钟₂供体和 DNIC 的形成与硫代硫酸盐配体，所有生物系统的通用 NO 信号分子，在细胞中。获得的结果扩展了对亚硝阳离子功能的认识，并有助于揭示 DNIC 供体的毒性活性机制。它们与我们早期在 DNICglu 实验中关于 NO ⁺抑制大肠杆菌菌落形成的结果以及其他研究人员在酵母和 MCF7 癌细胞培养物中获得的数据相关联。