Real-time and specific monitoring of nitric oxide and evaluating of the oxidative stress in living cells and zebrafish under the pollutant exposure using a carbon dot-based composite fluorescent probe

Abstract

Nitric oxide (NO) functions as an essential signalling molecule in various physiological and pathological pathways. In vitro and vivo redox processes mediated by reactive oxygen species (ROS) and nitric oxide (NO) directly influence the intracellular state. In this study, a red-emitting fluorescent nanoprobe, N,S-CDs@Zn-ICA, was synthesized to monitor NO fluctuations in living cells and zebrafish under the exposure to various pollutants. Red-emissive carbon dots (N,S-CDs) were synthesized by a hydrothermal method using o-phenylenediamine and urea as carbon / nitrogen sources, and H₂SO₄ as sulfur source. Glutathione (GSH) was introduced to link N,S-CDs with metal organic complexes (Zn-ICA) through an amidation reaction to fabricate a carbon dot-based composite fluorescent probe, which greatly improved the selectivity, stability, and response time of the N,S-CDs. The composite probe has high selectivity and sensitivity with limit of detection (LOD) of 96.0 nM. Furthermore, the proposed probe was successfully used to monitor the dynamic changes in NO levels and evaluate oxidative stress in MCF-7 cells and zebrafish under the exposure to various pollutants, including seven heavy metal ions (such as Pb²⁺, Cd²⁺, and Hg²⁺) and nine organic pollutants at different concentrations and exposure times. This work provides a novel strategy for constructing highly selective and red-emitting fluorescent probe for real-time and dynamic monitoring of NO and further evaluating oxidative stress induced by pollutants in vitro and in vivo via fluorescence imaging.

Keywords: Carbon dots; Fluorescence imaging; Nitric oxide; Oxidative stress; ROS.