Regular ArticleA Fluorophore-Containing Nitroxide as a Probe to Detect Superoxide and Hydroxyl Radical Generated by Stimulated Neutrophils
Abstract
Toward the development of a fluorescence assay in combination with confocal microscopy to image free radicals generated by cells, we synthesized a fluorophore-nitroxide, 5-((2-carboxy)phenyl)-5-hydroxy-1-((2,2,5,5-tetramethyl-1-oxypyrrolidin-3-yl)methyl)-3-phenyl-2-pyrrolin-4-one sodium salt, and tested the applicability of this probe to detect oxygen-centered free radicals. The reaction of the fluorophore-nitroxide with superoxide (10 μM/min) generated either by the reaction of xanthine oxidase on xanthine or by PMA-activated neutrophils in the presence of cysteine (200 μM) resulted in a loss of electron spin resonance (ESR) signal intensity concurrent with an increase in fluorescence emission. The decrease in ESR signal and the augmentation in fluorescence emission were inhibited by the addition of superoxide dismuatse. This fluorophore-nitroxide also reacted with methyl radical generated by the reaction of hydroxyl radical with DMSO (0.14 M). In this case a loss in ESR signal intensity concomitant with an increase in fluorescence emission which were inhibited by catalase (300 U/ml), was recorded. These results clearly demonstrated the feasibility of using fluorescence methodology in conjunction with a fluorophore-nitroxide to detect oxygen-centered free radicals in biological systems.
References (0)
Cited by (88)
Recent process in organic small molecular fluorescent probes for tracking markers of tumor redox balance
2024, TrAC - Trends in Analytical ChemistryExploring the redox balance in tumor region is of great significance for clarifying the pathogenesis of tumors and finding methods for the treatment of tumors. Tumors can maintain specific redox balance by expression of some types of reactive oxygen species (for instance O2•−, H2O2, ONOO−, etc), and reducing substances (GSH, H2Sn, Cys). These substances can thus be used as markers to identify the changes in redox balance in cancer cells or tumor microenvironments. However, their high reactivity makes them difficult to be rapidly detected in-situ in biosystems. To solve this problem, various types of organic small molecular fluorescent probes (OSMFPs) have been developed relying on fluorescence imaging technology. In this review, we have systematically introduced the recent progress about OSMFPs that could detect tumor redox balance and proposed some outlooks based on our opinions. This review provide important reference for further development of OSMFPs-based detection methods for redox balance.
Constructing fluorescence probes with fluoride-substituted donors for detecting •OH in living cells and zebrafish
2023, Dyes and PigmentsHydroxyl radicals (•OH) is one of the commonly reactive oxygen species available in biosystem and plays a key role in a number of physiological and pathological processes. Based on our previous study, fluoride could serve as a moderate electron donating group in constructing fluorescence probes. However, there is seldom work on the effects of fluoride-substituted moiety towards the photophysical properties in constructing fluorescence probes. In this work, we developed a series of novel fluorescence probes based on a quinolinium-scaffold containing fluoride substituted donor. From our results, the probe FHyR-3 which has an extended conjugated system could detect hydroxyl radical with high selectivity, sensitivity, and fast response. Furthermore, the probe has the maximum emission at 508 nm with a large stoke shift (114 nm) and exhibits low toxicity, good biocompatibility and mito-targeting. We also demonstrated its capability in tracking hydroxyl radical level in living cells and zebrafish.
Reactive Oxygen Species (ROS) and Reactive Nitrogen Species (RNS) in plants– maintenance of structural individuality and functional blend
2022, Advances in Redox ResearchThe free radicals along with several reactive oxygen species (ROS) and reactive nitrogen species (RNS) are known to play a dual role in the biological living system which carries a substantial importance in terms of signal networking in plants. The production of these active molecules in different cellular compartments eventually led to oxidative damage. However recent discoveries have evidenced its crucial roles as signaling molecules, activating stress responses against environmental challenges. As can be seen, the cellular organelles are considered to be the primary repository and site of action for reactive species there by later with the establishment of stress signaling concept, the underlying mechanism of ROS/RNS interaction has been elucidated properly by cellular organelle based study. These efforts led to the identification of signaling cascades generated by ROS and RNS which are not only involved with various antioxidative pathways but also correspond with other stress specific mechanisms. This study focuses on a burgeoning area of plant study, highlights the site specific generation, interplay, effect on several metabolic pathways and mode of reaction of ROS/RNS in cells. The review moreover postulates the fundamental mechanism of ROS/RNS cross talking in a lucid manner which further helps to stand out its significant importance with respect to plant survival during the course of evolution. Increasing interest in the area of plant stress and the ROS/RNS signaling, more elementary knowledge regarding their specificity, regulation, flexibility yet to be explored at molecular level by the advancement of technology.
Fluorescent probes for the detection of nitroxyl (HNO)
2018, Free Radical Biology and MedicineNitroxyl (HNO), which according to the IUPAC recommended nomenclature should be named azanone, is the protonated one-electron reduction product of nitric oxide. Recently, it has gained a considerable attention due to the interesting pharmacological effects of its donors. Although there has been great progress in the understanding of HNO chemistry and chemical biology, it still remains the most elusive reactive nitrogen species, and its selective detection is a real challenge. The development of reliable methodologies for the direct detection of azanone is essential for the understanding of important signaling properties of this reactive intermediate and its pharmacological potential. Over the last decade, there has been considerable progress in the development of low-molecular-weight fluorogenic probes for the detection of HNO, and therefore, in this review, we have focused on the challenges and limitations of and perspectives on nitroxyl detection based on the use of such probes.
Sensitive determination of endogenous hydroxyl radical in live cell by a BODIPY based fluorescent probe
2017, TalantaThe sensitive and selective fluorescence probe for hydroxyl radical analysis is of significance because hydroxyl radical plays key roles in many physiological and pathological processes. In this work, a novel organic fluorescence molecular probe OHP for hydroxyl radical is synthesized by a two-step route. The probe employs 4-bora-3a,4a-diaza-s-indacene (difluoroboron dipyrromethene, BODIPY) as the fluorophore and possesses relatively high fluorescence quantum yields (77.14%). Hydroxyl radical can rapidly react with the probe and quench the fluorescence in a good linear relationship (R2=0.9967). The limit of detection is determined to be as low as 11 nM. In addition, it has been demonstrated that the probe has a good stability against pH and light illumination, low cytotoxicity and high biocompatibility. Cell culture experimental results show that the probe OHP is sensitive and selective for imaging and tracking endogenous hydroxyl radical in live cells.
Reactive oxygen species: Reactions and detection from photosynthetic tissues
2015, Journal of Photochemistry and Photobiology B: BiologyReactive oxygen species (ROS) have long been recognized as compounds with dual roles. They cause cellular damage by reacting with biomolecules but they also function as agents of cellular signaling. Several different oxygen-containing compounds are classified as ROS because they react, at least with certain partners, more rapidly than ground-state molecular oxygen or because they are known to have biological effects. The present review describes the typical reactions of the most important ROS. The reactions are the basis for both the detection methods and for prediction of reactions between ROS and biomolecules. Chemical and physical methods used for detection, visualization and quantification of ROS from plants, algae and cyanobacteria will be reviewed. The main focus will be on photosynthetic tissues, and limitations of the methods will be discussed.