A novel activatable AIEgen fluorescent probe for peroxynitrite detection and its application in EC1 cells

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Highlights

  • It is the first QM-based AIE fluorescent probe for ONOO−.

  • QM-ONOO− showed bigger Stokes shift (190 nm) than the reported.

  • QM-ONOO− possessed rapid response, and low detection limit (27.5 nM).

  • The exogenous and endogenous ONOO− imaging in MCF-7 cells demonstrated its value of practical tool.

Abstract

Peroxynitrite (ONOO), as one type of reactive nitrogen species (RNS), is closely related to multiple pathophysiological processes. Visualization of ONOO in living system is essential for further study of its physiological functions. However, most existing ONOO fluorescent probes continuously display the aggregation-caused quenching (ACQ). In present research, a new activatable AIEgen fluorescent probe QM-ONOO for ONOO recognition and imaging was proposed. Activation of the probe by ONOO, induced aggregation of QM−OH. This sensor showed fast response time (<4 min) and high sensitivity (DL, 27.5 nM), large Stock's shift (190 nm), long emission wavelength. Furthermore, the outcomes indicate that QM-ONOO as an activatable AIEgen fluorescent sensor was robust chemical tool to detect endogenous peroxynitrite in living biological system.

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 (O2radical dot) 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)

  • B. Zhu et al.

    A highly selective and ultrasensitive ratiometric far-red fluorescent probe for imaging endogenous peroxynitrite in living cells

    Sens. Actuators B Chem.

    (2018)
  • P. Pacher et al.

    Nitric oxide and peroxynitrite in health and disease

    Physiol. Rev.

    (2007)
  • G. Ferrer-Sueta et al.

    Biochemistry of peroxynitrite and protein tyrosine nitration

    Chem. Rev.

    (2018)
  • R.T. Dean et al.

    Biochemistry and pathology of radical-mediated protein oxidation

    Biochem. J.

    (1997)
  • G.Y. Liou et al.

    Reactive oxygen species in cancer

    Free Radic. Res.

    (2010)
  • L. Myatt et al.

    Brockman D: oxidative stress causes vascular dysfunction in the placenta

    J. Matern. Med.

    (2000)
  • Z. Li et al.

    A novel dual-channel fluorescent probe for nitroxyl detection and its application in HepG-2 cells

    Sens. Actuators B Chem.

    (2020)
  • H. Niu et al.

    Mitochondria-targeted fluorescent probes for oxidative stress imaging

    Sens. Actuators B Chem.

    (2019)
  • X. Yang et al.

    A multi-signal mitochondria-targeted fluorescent probe for real-time visualization of cysteine metabolism in living cells and animals

    Chem. Commun. (Camb.)

    (2018)
  • Y. Chu et al.

    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.

    (2019)
  • C. Xu et al.

    A facile AIEgen-based fluorescent probe design strategy and its application in hypochlorite probe construction

    Sens. Actuators B Chem.

    (2020)
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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.

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