A novel activatable AIEgen fluorescent probe for peroxynitrite detection and its application in EC1 cells
Introduction
The peroxynitrite anion (ONOO−) has received special attention as a reactive nitrogen species (RNS), which is obtained via the diffusion-controlled chemical reaction of (O2−) and (NO) [1,2]. The anion can react in out of control manner with multiple small biological molecules owing to its powerful oxidability and nitrating characteristic [[3], [4], [5]]. The abnormal cumulation of peroxynitrite anion in living cells was certificated to be pernicious. It damages many critical small biological molecules such as proteins even nucleic acids, therefore affecting normal physiological function of cells, which even more cause cell necrosis and apoptosis. The mis-regulation of ONOO− in biological systems could result in Alzheimer's disease, inflammatory conditions, cardiovascular disease and cancer [[6], [7], [8], [9], [10]]. Consequently, to explore powerful tools to selectively visualize ONOO− is important for revealing its precise biological function and better diagnosis of related diseases.
The small molecule fluorescent imaging has aroused great interest, their possess obvious advantages in excellent biocompatibility, rapid detection, high selectivity, high sensitivity, fantastic temporal, spatial resolution and high safety [[11], [12], [13], [14], [15], [16], [17]]. To date, some fluorescent probes have been reported to monitor and visualizate ONOO− in cells (Table S1) [[18], [19], [20], [21], [22], [23], [24], [25], [26], [27], [28], [29], [30], [31], [32], [33], [34], [35]]. However, currently conventional fluorescent probes have hydrophobic feature with planar structure. Massive probes display aggregation-caused quenching (ACQ) problem in concentrated solution or aggregated state which lead to their emission is attenuate or even quenched. To the opposite common ACQ dyes, aggregation-induced emission (AIE) fluorescent dyes have rotatable structures or twisted propeller-shaped conformations, these are highly emissive because of the constraint of intramolecular movement. In particular, the higher concentrations, the stronger their emission become [36,37]. In consequence, AIE fluorophores are the ideal candidate for ONOO− detecting in biomolecules. Furthermore, probes with long emission wavelength (>600 nm) have showed little light damage, minimizes unnecessary influences of background fluorescence and deeper tissue penetration [38]. However, to our best knowledge, an activatable AIEgen probe with long emission wavelength detecting ONOO− has scarcely been reported.
The quinoline-malononitrile (QM) is good chromophore that contains several excellent features, such as the red emission, marked photostability, high brightness, good biocompatibility [37]. Mounting studies have shown that fluorescent probes containing arylboronic acid pinacol esters might higher selectivity and easily react towards ONOO− than other ROS [[39], [40], [41]]. In addition, arylboronates take precedence over reaction with ONOO− instead of H2O2 [38], although a variety of containing arylboronic acid pinacol esters fluorescent probes for detecting H2O2 have been reported [42]. Therefore, we designed a new QM-based long emission wavelength fluorescent probe QM-ONOO− for endogenous and exogenous ONOO− detection (shown in Scheme 1). The probe QM-ONOO− was consisted of QM chromophore and arylboronates group for ONOO− recognition. As expected, probe QM-ONOO− exhibits excellent sensing performance at high selectivity and sensitivity, long emission wavelength (λem =620 nm), low limit detection (27.5 nM) and fast response. The activatable AIEgen probe QM-ONOO− also showed bigger Stock's shift (190 nm) than the reported (Table S1). Upon the addition of ONOO−, QM-OH can be released. Then QM-OH can form fluorescent aggregates at the activation site which restricted free molecular rotation and restored its intrinsic AIE fluorescence. It accompany with a remarkable light-up fluorescent signal. Benefiting from these advantages, the long emission wavelength activatable AIEgen probe can provide a novel approach to track the fluctuation of ONOO− in living biological system.
Section snippets
Apparatus
Absorption spectra dates were tested by HP-8453 UV/Vis spectrometer. HITACHI F-4600 fluorescence spectrophotometer gathered fluorescent date. The excitation and emission slit was 5 nm, excitation voltage was 700 V. Dynamic light scattering (DLS) results were obtained on a NanoPlus-3 DLS particle size/zeta potential analyzer. Model pHs-3C meter was used to measure the pH. Bruker DTX-400 spectrometers were measured 1H and 13C NMR spectra in CDCl3 referencing tetramethylsilane (TMS). ESI mass
structure representation
QM-ONOO− was handily acquired by condensation reaction quinoline- malononitrile with l-1. Probe QM-ONOO− exactly features by 1H NMR, 13C NMR, and HR-MS spectrometry (Figs. S2 - S4.).
Photophysical properties QM−OH and probe QM-ONOO−
QM-ONOO− and QM−OH in the DMSO/PBS mixture are explored under excitation at 430 nm in the fluorescence spectra. We observed obvious difference. The fluorescence of QM-ONOO- always unchanged with water increasing (Fig. S5). The fluorescence of QM−OH will change as water increasing. When the water fraction increased
Conclusion
To conclude, a unique activatable AIEgen fluorescent probe QM-ONOO− was proposed to achieve ONOO− detection in live EC1 cells. Activatable AIEgen probe QM-ONOO− displayed long emission wavelength (620 nm), large Stoke's shift (∼190 nm), fast responsive recognition of ONOO−. Furthermore, experimental results indicated that QM-ONOO− could successfully discover exogenous and endogenous peroxynitrite in EC1 cells. Therefore, possessed great quantity outstanding properties, this seminal work can be
CRediT authorship contribution statement
Xiaojing Han: Data curation, Writing - original draft. Xiaopeng Yang: Methodology, Investigation. Yongru Zhang: Visualization. Zipeng Li: Software, Data curation. Wenbo Cao: Writing - review & editing, Validation. Di Zhang: Supervision. Yong Ye: Resources, Funding acquisition, Project administration, Supervision, Writing - review & editing.
Declaration of Competing Interest
None.
Acknowledgments
This work was financially supported by the National Science Foundation of China (No. 21572209 and 21907025), Scientific and Technological Project of Henan Province of China (192102310140), Key scientific research projects of colleges and universities in Henan Province (No. 18A310035).
Xiaojing Han is getting her MS degree at the College of Chemistry, Zhengzhou University. Her research interests focus on fluorescent sensors.
References (43)
- et al.
Evidence for nitric oxide-mediated oxidative damage in chronic inflammation. Nitrotyrosine in serum and synovial fluid from rheumatoid patients
FEBS Lett.
(1994) - et al.
Mass spectrometric identification of tryptophan nitration sites on proteins in peroxynitrite-treated lysates from PC12 cells
Free Radic. Biol. Med.
(2011) - et al.
Activation pattern of mitogen-activated protein kinases elicited by peroxynitrite: attenuation by selenite supplementation
FEBS Lett.
(1999) - et al.
A dual-channel probe with green and near-infrared fluorescence changes for in vitro and in vivo detection of peroxynitrite
Anal. Chim. Acta
(2019) - et al.
A rapid responsive colorimetric and near-infrared fluorescent turn-on probe for imaging exogenous and endogenous peroxynitrite in living cells
Sens. Actuators B Chem.
(2018) - et al.
A lysosome-targetable fluorescent probe for imaging ONOO− in living cells and animals
Dye. Pigment.
(2019) - et al.
A BODIPY fluorescence probe modulated by selenoxide spirocyclization reaction for peroxynitrite detection and imaging in living cells
Dye. Pigment.
(2013) - et al.
A novel hepatoma-specific fluorescent probe for imaging endogenous peroxynitrite in live HepG2 cells
Sens. Actuators B Chem.
(2019) - et al.
Reaction-based Indicator displacement Assay (RIA) for the selective colorimetric and fluorometric detection of peroxynitrite
Chem. Sci.
(2015) - et al.
A boronate-based fluorescent probe for the selective detection of cellular peroxynitrite
Chem. Commun. (Camb.)
(2014)
A highly selective and ultrasensitive ratiometric far-red fluorescent probe for imaging endogenous peroxynitrite in living cells
Sens. Actuators B Chem.
Nitric oxide and peroxynitrite in health and disease
Physiol. Rev.
Biochemistry of peroxynitrite and protein tyrosine nitration
Chem. Rev.
Biochemistry and pathology of radical-mediated protein oxidation
Biochem. J.
Reactive oxygen species in cancer
Free Radic. Res.
Brockman D: oxidative stress causes vascular dysfunction in the placenta
J. Matern. Med.
A novel dual-channel fluorescent probe for nitroxyl detection and its application in HepG-2 cells
Sens. Actuators B Chem.
Mitochondria-targeted fluorescent probes for oxidative stress imaging
Sens. Actuators B Chem.
A multi-signal mitochondria-targeted fluorescent probe for real-time visualization of cysteine metabolism in living cells and animals
Chem. Commun. (Camb.)
An Intramolecular Charge Transfer and Aggregation Induced EmissionEnhancement Fluorescent Probe Based on 2-Phenyl-1,2,3-triazole forHighly Selective and Sensitive Detection of Homocysteine andIts Application in Living Cells
Chin. J. Chem.
A facile AIEgen-based fluorescent probe design strategy and its application in hypochlorite probe construction
Sens. Actuators B Chem.
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Xiaojing Han is getting her MS degree at the College of Chemistry, Zhengzhou University. Her research interests focus on fluorescent sensors.
Xiaopeng Yang is getting his PhD degree at the College of Chemistry, Zhengzhou University. His current research interests focus on developing fluorescent probes.
Yongru Zhang is getting her MS degree at the College of Chemistry, Zhengzhou University. Her research interests focus on fluorescent sensors.
Zipeng Li is getting his Master degree at the College of Chemistry, Zhengzhou University. His current research interests focus on developing fluorescent probes.
Wenbo Cao is working at Zhengzhou University. His research interests focus on fluorescent sensors.
Di Zhang is now working in Institute of Agricultural Quality Standards and Testing Technology, Henan Academy of Agricultural Sciences. His current research interests include probes, chemical biology science.
Yong Ye received his PhD degree in 2003 from Nankai University, PR China. He is a professor at the College of Chemistry, Zhengzhou University. His current research interests including synthesis and sensors.